Use of il-1beta binding antibodies

ABSTRACT

Use of an IL-Iβ binding antibody or a functional fragment thereof, especially canakinumab or a functional fragment thereof, or gevokizumab or a functional fragment thereof, and biomarkers for the treatment of cancer with at least partial inflammatory basis, e.g., CML.

TECHNICAL FIELD

The present invention relates to the use of an IL-1β binding antibody ora functional fragment thereof, for the treatment of cancer, e.g., cancerhaving at least a partial inflammatory basis, e.g., CML.

BACKGROUND OF THE DISCLOSURE

Chronic myelogenous leukemia (CML, also known as Chronic myeloidleukemia) is a cancer of the bone marrow characterized by increased andunregulated clonal proliferation of predominantly myeloid cells in thebone marrow. Its annual incidence is 1-2 per 100,000 people, affectingslightly more men than women. CML represents about 15-20% of all casesof adult leukemia in Western populations, about 4,500 new cases per yearin the U.S. or in Europe (Faderl et al., N. Engl. J. Med. 1999, 341:164-72).

In CML a reciprocally balanced chromosomal translocation inhematopoietic stem cells (HSCs) produces the BCR-ABL hybrid gene. Thelatter encodes the oncogenic Bcr-Abl fusion protein. Whereas ABL encodesa tightly regulated protein tyrosine kinase, which plays a fundamentalrole in regulating cell proliferation, adherence and apoptosis, theBCR-ABL fusion gene encodes as constitutively activated kinase, whichtransforms HSCs to produce a phenotype exhibiting deregulated clonalproliferation, reduced capacity to adhere to the bone marrow stroma anda reduces apoptotic response to mutagenic stimuli, which enable it toaccumulate progressively more malignant transformations. The resultinggranulocytes fail to develop into mature lymphocytes and are releasedinto the circulation, leading to a deficiency in the mature cells andincreased susceptibility to infection.

Imatinib mesylate (STI571, GLEEVEC®) is the standard of therapy for CMLwith response rates of more than 96%, and works by inhibiting theactivity of BCR-ABL. However, despite initial success, patientseventually develop resistance to imatinib mesylate due to acquisition ofpoint mutations in BCR-ABL. Therefore, there remains a continued need todevelop new treatment options for CML.

SUMMARY OF THE DISCLOSURE

The present disclosure relates to the use of an IL-1β binding antibodyor a functional fragment thereof, suitably canakinumab, suitablygevokizumab, for the treatment of cancers, e.g., cancers that have atleast a partial inflammatory basis, e.g., CML. In another aspect, thepresent invention relates to a particular clinical dosage regimen forthe administration of an IL-1β binding antibody or a functional fragmentthereof for the treatment of cancers, e.g., cancers that have at least apartial inflammatory basis, e.g., CML. In one embodiment the preferreddose of canakinumab for a patient with cancer that has at least apartial inflammatory basis, e.g., CML, is about 200 mg about every 3weeks or about monthly, preferably subcutaneously. In one embodiment thepatient receives gevokizumab about 30 mg to about 120 mg per treatmentabout every 3 weeks or about monthly, preferably intravenously. Inanother aspect the subject with cancer having at least a partialinflammatory basis, e.g., CML, is administered with one or moretherapeutic agent (e.g., a chemotherapeutic agent) and/or hasreceived/will receive debulking procedures in addition to theadministration of an IL-1β binding antibody or a functional fragmentthereof.

There are also provided methods of treatment of cancers that have atleast a partial inflammatory basis, e.g., CML, in a patient in needthereof comprising administering to the subject a therapeuticallyeffective amount of an IL-1β binding antibody or a functional fragmentthereof.

Another aspect of the invention is the use of an IL-1β binding antibodyor a functional fragment thereof, for the preparation of a medicamentfor the treatment of cancer having at least a partial inflammatorybasis, e.g., CML.

The present disclosure also provides a pharmaceutical compositioncomprising a therapeutically effective amount of an IL-1β bindingantibody or a functional fragment thereof, suitably canakinumab orgevokizumab, for use in the treatment of cancer having at least apartial inflammatory basis, e.g., CML, in a patient. In one embodiment,the pharmaceutical composition comprising a therapeutically effectiveamount of an IL-1β binding antibody or a functional fragment thereof,e.g., canakinumab, is in the form of an autoinjector. In one embodimentabout 200 mg of canakinumab is loaded in an autoinjector.

In one aspect the present invention provides an IL-1β binding antibodyor a functional fragment thereof (e.g., canakinumab or gevokizumab), foruse in a patient in need thereof in the treatment of a cancer, e.g., acancer having at least partial inflammatory basis, e.g., CML. Each andevery embodiments disclosed in this application applies, separately orin combination, to this aspect.

In one aspect the present invention provides an IL-1β binding antibodyor a functional fragment thereof (e.g., canakinumab or gevokizumab), foruse in a patient in need thereof in the treatment of a cancer having atleast partial inflammatory basis, e.g., CML. Each and every embodimentsdisclosed in this application applies, separately or in combination, tothis aspect.

In one aspect the present invention provides an IL-1β binding antibodyor a functional fragment thereof (e.g., canakinumab or gevokizumab), foruse in a patient in need thereof in the treatment of CML.

FIGURE LEGENDS

FIG. 1. In vivo model of spontaneous human breast cancer metastasis tohuman bone predicts a key role for IL-1β signaling in breast cancer bonemetastasis. Two 0.5 cm³ pieces of human femoral bone were implantedsubcutaneously into 8-week old female NOD SCID mice (n=10/group). 4weeks later luciferase labelled MDA-MB-231-luc2-TdTomato or T47D cellswere injected into the hind mammary fat pads. Each experiment wascarried out 3-separate times using bone form a different patient foreach repeat. Histograms showing fold change of IL-1B, IL-1R1, Caspase 1and IL-1Ra copy number (dCT) compared with GAPDH in tumour cells grownin vivo compared with those grown in a tissue culture flask (a i);mammary tumours that metastasize compared with mammary tumours tumoursthat do not metastasize (a ii); circulating tumour cells compared withtumour cells that remain in the fat pad (a iii) and bone metastasescompared with the matched primary tumour (a iv). Fold change in IL-1βprotein expression is shown in (b) and fold change in copy number ofgenes associated with EMT (E-cadherin, N-cadherin and JUP) compared withGAPDH are shown in (c). *=P<0.01, **=P<0.001, ***=P<0.0001, {circumflexover ( )}{circumflex over ( )}{circumflex over ( )}=P<0.001 comparedwith naïve bone.

FIG. 2. Stable transfection of breast cancer cells with IL-1B.MDA-MB-231, MCF7 and T47D breast cancer cells were stably transfectedwith IL-1B using a human cDNA ORF plasmid with a C-terminal GFP tag orcontrol plasmid. a) shows pg/ng IL-1β protein from IL-1β-positive tumourcell lysates compared with scramble sequence control. b) shows pg/ml ofsecreted IL-1β from 10,000 IL-1β+ and control cells as measured byELISA. Effects of IL-1B overexpression on proliferation of MDA-MB-231and MCF7 cells are shown in (c and d) respectively. Data shown aremean+/−SEM, *=P<0.01, **=P<0.001, ***=P<0.0001 compared with scramblesequence control.

FIG. 3. Tumour derived IL-1β induces epithelial to mesenchymaltransition in vitro. MDA-MB-231, MCF7 and T47D cells were stablytransfected with to express high levels of IL-1B, or scramble sequence(control) to assess effects of endogenous IL-1B on parameters associatedwith metastasis. Increased endogenous IL-1B resulted tumour cellschanging from an epithelial to mesenchymal phenotype (a). b) showsfold-change in copy number and protein expression of IL-1B, IL-1R1,E-cadherin, N-cadherin and JUP compared with GAPDH and β-cateninrespectively. Ability of tumour cells to invade towards osteoblaststhrough Matrigel and/or 8 μM pores, are shown in (c) and capacity ofcells to migrate over 24 and 48 h is shown using a wound closure assay(d). Data are shown as mean+/−SEM, *=P<0.01, **=P<0.001, ***=P<0.0001.

FIG. 4. Pharmacological blockade of IL-1β inhibits spontaneousmetastasis to human bone in vivo. Female NOD-SCID mice bearing two 0.5cm³ pieces of human femoral bone received intra-mammary injections ofMDA-MB-231Luc2-TdTomato cells. One week after tumour cell injection micewere treated with 1 mg/kg/day IL-1Ra, 20 mg/kg/14-days canakinumab, orplacebo (control) (n=10/group). All animals were culled 35 daysfollowing tumour cell injection. Effects on bone metastases (a) wereassessed in vivo and immediately post-mortem by luciferase imaging andconfirmed ex vivo on histological sections. Data are shown as numbers ofphotons per second emitted 2 minutes following sub-cutaneous injectionof D-luciferin. Effects on numbers of tumour cells detected in thecirculation are shown in (b). *=P<0.01, **=P<0.001, ***=P<0.0001.

FIG. 5. Tumour derived IL-1β promotes breast cancer bone homing in vivo.8-week old female BALB/c nude mice were injected with control (scramblesequence) or IL-1β overexpressing MDA-MB-231-IL-1β+ cells via thelateral tail vein. Tumour growth in bone and lung were measured in vivoby GFP imaging and findings confirmed ex vivo on histological sections.a) shows tumour growth in bone; b) shows representative μCT images oftumour bearing tibiae and the graph shows bone volume (BV)/tissue volume(TV) ratio indicating effects on tumour induced bone destruction; c)shows numbers and size of tumours detected in lungs from each of thecell lines. *=P<0.01, **=P<0.001, ***=P<0.0001. (B=bone, T=tumour,L=lung)

FIG. 6. Tumour cell-bone cell interactions stimulate IL-1β productioncell proliferation. MDA-MB-231 or T47D human breast cancer cell lineswere cultured alone or in combination with live human bone, HS5 bonemarrow cells or OB1 primary osteoblasts. a) shows the effects ofculturing MDA-MB-231 or T47D cells in live human bone discs on IL-1βconcentrations secreted into the media. The effect of co-culturingMDA-MB-231 or T47D cells with HS5 bone cells on IL-1β derived from theindividual cell types following cell sorting and the proliferation ofthese cells are shown in b) and c). Effects of co-culturing MDA-MB-231or T47D cells with OB1 (osteoblast) cells on proliferation are shown ind). Data are shown as mean+/−SEM, *=P<0.01, **=P<0.001, ***=P<0.0001.

FIG. 7. IL-1β in the bone microenvironment stimulates expansion of thebone metastatic niche. Effects of adding 40 pg/ml or 5 ng/ml recombinantIL-1β to MDA-MB-231 or T47D breast cancer cells is shown in (a) andeffects on adding 20 pg/ml, 40 pg/ml or 5 ng/ml IL-1B on proliferationof HS5, bone marrow, or OB1, osteoblasts, are shown in b) and c)respectively. (d) IL-1 driven alterations to the bone vasculature wasmeasured following CD34 staining in the trabecular region of the tibiaefrom 10-12-week old female IL-1R1 knockout mice. (e) BALB/c nude micetreated with 1 mg/ml/day IL-1Ra for 31 days and (f) C57BL/6 mice treatedwith 10 μM canakinumab for 4-96 h. Data are shown as mean+/−SEM,*=P<0.01, **=P<0.001, ***=P<0.0001.

FIG. 8. Suppression of IL-1 signalling affects bone integrity andvasculature. Tibiae and serum from mice that do not express IL-1R1(IL-1R1 KO), BALB/c nude mice treated daily with 1 mg/kg per day ofIL-1R antagonist for 21 and 31 days and C57BL/6 mice treated with 10mg/kg of canakinumab (Ilaris) of 0-96 h were analysed for bone integrityby μCT and vasculature using ELISA for Endothelin 1 and pan VEGF. a)shows the effects of IL-1R1 KO; b) effects of Anakinra and c) effects ofcanakinumab on bone volume compared with tissue volume (i),concentration of Endothelin 1 (ii) and concentrations of VEGF secretedinto the serum. Data shown are mean+/−SEM, *=P<0.01, **=P<0.001,***=P<0.0001 compared with control.

FIG. 9. Tumour derived IL-1β predicts future recurrence and bone relapsein patients with stage II and III breast cancer. ˜1300 primary breastcancer samples from patients with stage II and III breast cancer with noevidence of metastasis were stained for 17 kD active IL-1β. Tumours werescored for IL-1β in the tumour cell population. Data shown are KaplanMeyer curves representing the correlation between tumour derived IL-1βand subsequent recurrence a) at any site or b) in bone over a 10-yeartime period.

FIG. 10. Simulation of canakinumab PK profile and hsCRP profile. a)shows canakinumab concentration time profiles. Solid line and band:median of individual simulated concentrations with 2.5-97.5% predictioninterval (300 mg Q12W (bottom line), 200 mg Q3W (middle line), and 300mg Q4W (top line)). b) shows the proportion of month 3 hsCRP being belowthe cut point of 1.8 mg/L for three different populations: all CANTOSpatients (scenario 1), confirmed lung cancer patients (scenario 2), andadvanced lung cancer patients (scenario 3) and three different doseregimens. c) is similar to b) with the cut point being 2 mg/L. d) showsthe median hsCRP concentration over time for three different doses. e)shows the percent reduction from baseline hsCRP after a single dose.

FIG. 11. Gene expression analysis by RNA sequencing in colorectal cancerpatients receiving PDR001 in combination with canakinumab, PDR001 incombination with everolimus and PDR001 in combination with others. Inthe heatmap figure, each row represents the RNA levels for the labelledgene. Patient samples are delineated by the vertical lines., with thescreening (pre-treatment) sample in the left column, and the cycle 3(on-treatment) sample in the right column. The RNA levels arerow-standardized for each gene, with black denoting samples with higherRNA levels and white denoting samples with lower RNA levels.Neutrophil-specific genes FCGR3B, CXCR2, FFAR2, OSM, and G0S2 are boxed.

FIG. 12. Clinical data after gevokizumab treatment (panel a) and itsextrapolation to higher doses (panels b, c, and d). Adjusted percentchange from baseline in hsCRP in patients in a). The hsCRPexposure-response relationship is shown in b) for six different hsCRPbase line concentrations. The simulation of two different doses ofgevokizumab is shown in b) and c).

FIG. 13. Effect of anti-IL-1beta treatment in two mouse models ofcancer. a), b), and c) show data from the MC38 mouse model, and d) ande) show data from the LL2 mouse model.

FIG. 14. Efficacy of canakinumab in combination with pembrolizumab ininhibiting tumor growth.

FIG. 15. Preclinical data on the efficacy of canakinumab in combinationwith docetaxel in the treatment of cancer.

FIG. 16. Mice were implanted with 4T1 cells sc and treated with theindicated treatments on days 8 and 15 post tumor implant. There were 10mice in each group.

FIG. 17. Neutrophils (top) and monocytes (bottom) in 4T1 tumors 5 daysafter a single dose of docetaxel, 01BSUR, or the combination ofdocetaxel and 01BSUR.

FIG. 18. Granulocytic (top) and monocytic (bottom) MDSC in 4T1 tumors 5days after a single dose of docetaxel, 01BSUR, or the combination ofdocetaxel and 01BSUR.

FIG. 19. TIM-3+ CD4⁺ (top) and CD8⁺ (bottom) T cells in 4T1 tumors 4days after a second dose of docetaxel, 01BSUR, or the combination ofdocetaxel and 01BSUR.

FIG. 20. TIM-3 expressing Tregs in 4T1 tumors 4 days after a second doseof docetaxel, 01BSUR, or the combination of docetaxel and 01BSUR.

DETAILED DESCRIPTION OF THE DISCLOSURE

Many malignancies arise in areas of chronic inflammation and inadequateresolution of inflammation is hypothesized to play a major role in tumorinvasion, progression, and metastases (Voronov E, et al, PNAS 2003). Inmurine models, inflammasome activation and IL-1β production canaccelerate tumor invasiveness, growth, and metastatic spread (Voronov E,et al, PNAS 2003). For example, in IL-1β−/− mice, neither local tumorsnor lung metastases develop following localized or intravenousinoculation with melanoma cell lines, data suggesting that IL-1β may beessential for the invasiveness of already existing malignancies. It hasthus been hypothesized that inhibition of IL-1β might have an adjunctiverole in the treatment of cancers that have at least a partialinflammatory basis.

Advanced studies in delineating interaction between tumor and the tumormicroenvironment have revealed that chronic inflammation can promotetumor development, and tumor fuels inflammation to facilitate tumorprogression and metastasis. Inflammatory microenvironment with cellularand non-cellular secreted factors provides a sanctuary for tumorprogression by inducing angiogenesis; recruiting tumor promoting, immunesuppressive cells and inhibiting immune effector cell mediatedanti-tumor immune response. One of the major inflammatory pathwayssupporting tumor development and progression is IL-1β, apro-inflammatory cytokine produced by tumor and tumor associated immunesuppressive cells including neutrophils and macrophages in tumormicroenvironment.

In patients with myeloproliferative neoplasms (MPN), chronicinflammation has been identified as a potential initiating event as wellas a driver of clonal expansion that predisposes to a second cancer.High basal inflammatory status promotes mutagenesis through induction ofchronic oxidative stress and subsequent DNA oxidative damage, andelicits epigenetic changes that further promote inflammation(Hasselbalch 2013).

The MPN population has a significant inflammation-mediated comorbidityburden, ranging from second cancer to cardiovascular and thromboembolicdisease, chronic kidney disease, autoimmune disease and osteopenia(Hasselbalch 2015). One of the cytokine families most related to innateimmune responses and inflammation is the IL-1 family. IL-1β stands outas initiator of inflammatory processes; its role in hematologicalmalignancies has been described with promising therapeutic value showedin preclinical models (Arranz 2017).

IL-1β modulates hematopoietic stem cell (HSC) function. In preclinicalmodels, it promotes HSC differentiation biased into the myeloid linage.While acute IL-1β exposure contributes to HSC regeneration aftermyeloablation and transplantation chronic exposure promotes uncontrolledHSC division and exhaustion of the HSC pool (Pietras 2016). In contrast,inhibition of IL-1β signaling using IL-1Ra reduces colony formation exvivo (Zhang 2009). In vivo, IL-1Ra suppresses cell cycle in bone marrowHSC, and reduces leukocytes and platelets levels (Zhang 2009). Thus,preclinical models show that IL-1β levels play a physiological role inhematopoiesis, and suggest that their dysregulation participate inhematological diseases.

Philadelphia positive CML is classified as an MPN disorder. IncreasedIL-1β is seen in advanced blast phase as compared to chronic phase andhealthy controls, suggesting that is a marker of poor prognosis andshorter survival (Matti 2014). CML patients may display relapses throughmechanisms dependent on BCR-ABL or through additional mutations, likethose in genes promoting HSC survival or multidrug resistance.Importantly, IL-1β contributes to resistance to BCR-ABL tyrosine kinaseinhibitor imatinib in CML cells, where it increases cell survival anddecreases apoptosis rate through cyclooxygenase 2 (Lee 2016). Interferon(IFN) family members, alternative treatment against CML, haveanti-inflammatory effects and inhibit IL-1β. Higher levels of IL-1β wereseen in IFN-α-resistant CML patients as compared to sensitive patientsand healthy controls, and IL-1β stimulates colony growth inIFN-α-sensitive CML cells (Estrov 1991).

BCR-ABL tyrosine kinase inhibitors (TKI) are remarkably effective ininducing remissions and prolonging survival of CML patients. However,TKI treatment fails to eliminate CML leukemia stem cells (LSC), even inpatients achieving deep molecular responses (Chu 2011). Although asubset of CML patients are able to maintain remission after stoppingTKIs, most patients require continued treatment to prevent relapse. CMLLSC demonstrated increased expression of the IL-1 receptors, andenhanced sensitivity to IL-1-induced NF-KB signaling compared to normalstem cells. Treatment with recombinant IL-1 receptor antagonist (IL-1RA)inhibited IL-1 signaling in CML LSC and inhibited growth of CML LSC.Importantly, the combination of IL-1RA with nilotinib resulted insignificantly greater inhibition of CML LSC compared with TKI alone(Zhang 2015). Published literature also suggest that IL-1 signalingcontributes to overexpression of inflammatory mediators in CML LSC,suggesting that blocking IL-1 signaling could modulate the inflammatorymilieu. Zhang et al conclude that IL-1 signaling contributes tomaintenance of CML LSC following TKI treatment, and that IL-1 blockadewith IL-1RA enhances elimination of TKI-treated CML LSC. These resultsprovide a strong rationale for further exploration of anti-IL-1strategies to enhance LSC elimination in CML. IL-1 mediated signalingcontributes to resistance of CML LSC to TKI (Zhang 2015). Thecombination of TKIs with Il-1β inhibitors could be a strategy toincrease the proportion of CML patients achieving treatment-freeremission.

As reported in Rikder et all (Lancet, 2017), a randomised, double-blind,placebo-controlled trial of canakinumab in 10061 patients withatherosclerosis who had had a myocardial infarction, were free ofpreviously diagnosed cancer, and had concentrations of high-sensitivityC-reactive protein (hsCRP) of 2 mg/L or greater was completed in June,2017 (CANTOS trial). To assess dose-response effects, patients wererandomly assigned by computer-generated codes to three canakinumab doses(50 mg, 150 mg, and 300 mg, subcutaneously every 3 months) or placebo.

Baseline concentrations of hsCRP (median 6.0 mg/L vs 4.2 mg/L; p<0.0001)and interleukin 6 (3.2 vs 2.6 ng/L; p<0.0001) were significantly higheramong participants subsequently diagnosed with lung cancer than amongthose not diagnosed with cancer. During median follow-up of 3.7 years,compared with placebo, canakinumab was associated with dose-dependentreductions in concentrations of hsCRP of 26-41% and of interleukin 6 of25-43% (p<0.0001 for all comparisons). Total cancer mortality (n=196)was significantly lower in the pooled canakinumab group than in theplacebo group (p=0.0007 for trend across groups), but was significantlylower than placebo only in the 300 mg group individually (hazard ratio[HR] 0.49 [95% CI 0.31-0.75]; p=0.0009). Incident lung cancer (n=129)was significantly less frequent in the 150 mg (HR 0.61 [95% CI0.39-0.97]; p=0.034) and 300 mg groups (HR 0.33 [95% CI 0.18-0.59];p<0.0001; p<0.0001 for trend across groups). Lung cancer mortality wassignificantly less common in the canakinumab 300 mg group than in theplacebo group (HR 0.23 [95% CI 0.10-0.54]; p=0.0002) and in the pooledcanakinumab population than in the placebo group (p=0.0002 for trendacross groups).

Biomarker analysis of patients of non-lung cancers from the CANTOStrial, especially of the GI/GU cancers, has revealed that they haveelevated baseline hsCRP level and IL-6 level. In addition, GI/GU cancerpatients with higher baseline level of hsCRP and IL-6 seem to have ashorter time to cancer diagnosis than patients having lower baselinelevel (Example 11), suggesting the likelihood of the involvement ofIL-1β mediated inflammation in broader cancer indications, besides lungcancer, which warranties targeting IL-1β in the treatment of thosecancers. In addition hsCRP level and IL-6 level in GI/GU patients werereduced in the range comparable to other patients in the CANTOS trialtreatment group, suggesting inhibition of IL-1β signaling in thosepatients. Inhibition of IL-1β alone or preferably in combination withother anti-cancer agents could result in clinical benefit in treatingcancer, e.g., cancer having at least partial inflammatory basis, e.g.,CML, as further supported by data presented in the Examples 9-11.

Thus in one aspect, the present invention provides the use of an IL-1βbinding antibody or a functional fragment thereof (The term “an IL-1βbinding antibody or a functional fragment thereof” is sometimes referredas “DRUG of the invention” in this application, which should beunderstood as identical term), suitably canakinumab or a functionalfragment thereof (included in DRUG of the invention), gevokizumab or afunctional fragment thereof (included in DRUG of the invention), for thetreatment of cancers, e.g., cancers that have at least a partialinflammatory basis, e.g., CML. In a preferred embodiment, CML has atleast a partial inflammatory basis.

Accordingly, the present disclosure provides a method of treatingcancers that have at least a partial inflammatory basis, e.g., CML,using an IL-1β binding antibody or a functional fragment thereof. In apreferred embodiment, such IL-1β binding antibodies or functionalfragments thereof can reduce inflammation and/or improve the tumormicroenvironment, e.g., they can inhibit IL-1β mediated inflammation andIL-1β mediated immune suppression in the tumor microenvironment. Anexample of using an IL-1β binding antibody in modulating the tumormicroenvironment is shown in Example 7-9 herein. In some embodiments, anIL-1β binding antibody or a functional fragment thereof is used alone asa monotherapy. In some embodiments, an IL-1β binding antibody or afunctional fragment thereof is used in combination with another therapy,such as with a check point inhibitor or with one or morechemotherapeutic agent. As discussed herein, inflammation can promotetumor development, an IL-1β binding antibody or a functional fragmentthereof, either alone or in combination with another therapy, can beused to treat any cancer that can benefit from reduced IL-1β mediatedinflammation and/or improved tumor environment. An inflammationcomponent is universally present, albeit to different degrees, in thecancer development.

The meaning of “cancers that have at least a partial inflammatory basis”or “cancer having at least a partial inflammatory basis” is well knownin the art and as used herein refers to any cancer in which IL-1βmediated inflammatory responses contribute to tumor development and/orpropagation, including but not necessarily limited to metastasis. Suchcancer generally has concomitant inflammation activated or mediated inpart through activation of the Nod-like receptor protein 3 (NLRP3)inflammasome with consequent local production of interleukin-1β. In apatient with such cancer, the expression, or even the overexpression ofIL-1β can be generally detected, commonly at the site of the tumor,especially in the surrounding tissue of the tumor, in comparison tonormal tissue. The expression of IL-1β can be detected by routinemethods known in the art, such as immunostaining, ELISA-based assays,ISH, microarray, RNA sequencing or RT-PCR in the tumor as well as inserum/plasma. The expression or higher expression of IL-1β can beconcluded, for example, against a negative control, usually normaltissue at the same site. The expression or higher expression of IL-1βcan be also concluded, for example, when a higher than normal level ofIL-1β is found in serum/plasma. Simultaneously or alternatively, apatient with such cancer has generally chronic inflammation, which ismanifested, typically, by higher than normal level of CRP or hsCRP, IL-6or TNFα. Cancers, particularly cancers that have at least a partialinflammatory basis include but are not limited to CML. Cancers alsoinclude cancers that may not express IL-1β until after previoustreatment of such cancer, e.g., including treatment with achemotherapeutic agent, e.g., as described herein, which contribute tothe expression of IL-1β in the tumor and/or tumor microenvironment. Insome embodiments, the methods and uses comprise treating a patient thathas relapsed or a patient whose cancer is recurring after treatment withsuch agent. In other embodiments, the agent is associated with IL-1βexpression and the IL-1β antibody or functional fragment thereof isgiven in combination with such agent.

Inhibition of IL-1β resulted in reduced inflammation status, includingbut not limited to reduced hsCRP or IL-6 level. Thus the effect of thepresent invention in cancer patients can be measured by reducedinflammation status, including but not limited to reduced hsCRP or IL-6level.

The term “cancers that have at least a partial inflammatory basis” or“cancer having at least a partial inflammatory basis” also includescancers that benefit from the treatment of an IL-1β binding antibody ora functional fragment thereof. As inflammation in general contributes totumor growth at already an early stage, administration of an IL-1βbinding antibody or a functional fragment thereof (canakinumab orgevokizumab) could potentially stop tumor growth effectively at theearly stage or delay tumor progression effectively at the early stage,even though the inflammation status, such as expression oroverexpression of IL-1β, or the elevated level of CRP or hsCRP, IL-6 orTNFα, is still not apparent or measurable. However, patients havingearly stage CML still can benefit from the treatment of IL-1β bindingantibody or a functional fragment thereof, which can be manifested inclinical trials. The clinical benefit can be measured by, including butnot limited to disease-free survival (DFS), progression-free survival(PFS), overall response rate (ORR), disease control rate (DCR), durationof response (DOR) and overall survival (OS), preferably in a clinicaltrial setting, against placebo group or against effects achieved bystandard of care drugs. If a patient treated with the DRUG of theinvention has shown any improvement in one or more of the aboveparameters in comparison to the same treatment but without the DRUG ofthe invention, the patient is considered to have benefited from thetreatment.

Available techniques known to the skilled person in the art allowdetection and quantification of IL-1β in tissue as well as inserum/plasma, particularly when the IL-1β is expressed to a higher thannormal level. For example, using the R&D Systems high sensitivity IL-1βELISA kit, IL-1β cannot be detected in majority of healthy donor serumsamples, as shown in the following Table.

Sample Values

Serum/Plasma—Samples from apparently healthy volunteers were evaluatedfor the presence of human IL-1β in this assay. No medical histories wereavailable for the donors used in this study.

Mean of % Range Sample Type Detectable (pg/mL) Detecable (pg/mL) Serum(n = 50 0.357 10 ND-0.606 EDTA plasma (n = 50) 0.292 12 ND-0.580 Heparinplasma (n = 50) 0.448 14 ND-1.08 ND = Non-detectable

Thus in a healthy person the IL-1β level is barely detectable or justabove the detection limit according to this test with the highsensitivity R&D Systems IL-1β ELISA kit. It is expected that in apatient with cancer having at least partial inflammatory basis ingeneral has higher than normal level of IL-1β and can be detected by thesame kit. Taking the IL-1β expression level in a healthy person as thenormal level (reference level), the term “higher than normal level ofIL-1β” means an IL-1β level that is higher than the reference level.Normally at least about 2 fold, at least about 5 fold, at least about 10fold, at least about 15 fold, at least about 20 fold of the referencelevel is considered as higher than normal level. Blocking the IL-1βpathway normally triggers the compensating mechanism leading to moreproduction of IL-1β. Thus the term “higher than normal level of IL-1β”also means and includes the level of IL-1β either post, or morepreferably, prior to the administration of an IL-1β binding antibody ora fragment thereof. Treatment of cancer with agents other than IL-1βinhibitors, such as some chemotherapeutic agents, can result inproduction of IL-1β in the tumor microenvironment. Thus the term “higherthan normal level of IL-1β” also refers to the level of IL-1β eitherprior to or post to the administration of such an agent.

When using staining, such as immunostaining, to detect IL-1β expressionin a tissue preparation, the term “higher than normal level of IL-1β”means to that the staining signal generated by specific IL-1β protein orIL-1β RNA detecting molecule is distinguishably stronger than stainingsignal of the surrounding tissue not expressing IL-1β.

Available techniques are known to the skilled person in the art allowdetection and quantification of IL-6 in tissue as well as inserum/plasma, particularly when the IL-6 is expressed to a higher thannormal level. For example, using the R&D Systems (www.RnDsystems.com)“high quantikine HS ELISA, human IL-6 Immunoassay”, IL-6 can be detectedin majority of healthy donor serum samples, as shown in the followingTable.

It is expected that in a patient with cancer having at least partialinflammatory basis in general has higher than normal level of IL-6 andcan be detected by the same kit. Taking the IL-6 expression level in ahealthy person as the normal level (reference level), the term “higherthan normal level of IL-6” means an IL-6 level that is higher than thereference level, normally higher than about 1.9 pg/ml, higher than v2pg/ml, higher than about 2.2 pg/ml, higher than about 2.5 pg/ml, higherthan about 2.7 pg/ml, higher than about 3 pg/ml, higher than about 3.5pg/ml, or higher than about 4 pg/ml, as determined preferably by the R&Dkit mentioned above. Blocking the IL-1β pathway normally triggers thecompensating mechanism leading to more production of IL-1β. Thus theterm “higher than normal level of IL-6” also means and includes thelevel of IL-6 either post, or more preferably, prior to theadministration of an IL-1β binding antibody or a fragment thereof.Treatment of cancer with agents other than IL-1β inhibitors, such assome chemotherapeutic agents, can result in production of IL-1β in thetumor microenvironment. Thus the term “higher than normal level of IL-6”also refers to the level of IL-6 either prior to or post theadministration of such an agent.

When using staining, such as immunostaining, to detect IL-1β expressionin a tissue preparation, the term “higher than normal level of IL-6”means to that the staining signal generated by specific IL-6 protein orIL-6 RNA detecting molecule is distinguishably stronger than stainingsignal of the surrounding tissue not expressing IL-6.

As used herein, the terms “treat”, “treatment” and “treating” refer tothe reduction or amelioration of the progression, severity and/orduration of a disorder, e.g., a proliferative disorder, or theamelioration of one or more symptoms, suitably of one or morediscernible symptoms, of the disorder resulting from the administrationof one or more therapies. In specific embodiments, the terms “treat”,“treatment” and “treating” refer to the amelioration of at least onemeasurable physical parameter of a proliferative disorder, such asgrowth of a tumor, not necessarily discernible by the patient. In otherembodiments the terms “treat”, “treatment” and “treating” refer to theinhibition of the progression of a proliferative disorder, eitherphysically by, e.g., stabilization of a discernible symptom,physiologically by, e.g., stabilization of a physical parameter, orboth. In other embodiments the terms “treat”, “treatment” and “treating”refer to the reduction or stabilization of tumor size or cancerous cellcount. As far as cancers as discussed here, taking CML as an example,the term treatment refers to at least one of the following: alleviatingone or more symptoms of CML, delaying progression of CML, prolongingoverall survival, prolonging progression free survival, achieving MR4.5(a 4.5 log reduction of BCR-ABL/ABL transcript, i.e. BCR-ABL/ABL≤0.0032% IS (International Scale)) at 12 months, and TFR (treatment-freeremission) eligibility after 96 weeks of treatment.

In one embodiment, the patient is newly diagnosed with CML. In apreferred embodiment, CML is diagnosed in chronic phase (CML-CP). In apreferred embodiment, the patient the patient is diagnosed with CML-CPwith resistance, failure or intolerance to one prior TKI therapy.Failure is defined for CML-CP patients (CP at the time of initiation oflast therapy) as follows (patients must meet at least one of thefollowing criteria):

-   -   Three months after the initiation of therapy: No CHR (complete        hematologic response) or >95% Ph⁺ metaphases.    -   Six months after the initiation of therapy: BCR-ABL ratio >10%        IS and/or >65% Ph⁺ metaphases.    -   Twelve months after initiation of therapy: BCR-ABL ratio >10% IS        and/or >35% Ph⁺ metaphases.    -   At any time after the initiation of therapy, loss of CHR, CCyR        (complete cytogenetic response) or PCyR (partial cytogenetic        response).    -   At any time after the initiation of therapy, the development of        new BCR-ABL mutations.    -   At any time after the initiation of therapy, confirmed loss of        MMR (major molecular response) in 2 consecutive tests, of which        one must have a BCR-ABL ratio ≥1% IS.    -   At any time after the initiation of therapy, new clonal        chromosome abnormalities in Ph⁺ cells: CCA/Ph+.

Intolerance is defined as:

-   -   Non-hematologic intolerance: Patients with grade 3 or 4 toxicity        while on therapy, or with persistent grade 2 toxicity,        unresponsive to optimal management, including dose adjustments        (unless dose reduction is not considered in the best interest of        the patient if response is already suboptimal).    -   Hematologic intolerance: Patients with grade 3 or 4 toxicity        (absolute neutrophil count [ANC] or platelets) while on therapy        that is recurrent after dose reduction to the lowest doses        recommended by manufacturer.

Preferably, the patient has not been previously treated with ahematopoietic stem-cell transplantation, and the patient does not planto undergo allogeneic hematopoietic stem cell transplantation.Preferably, the patient has not had Cardiac or cardiac repolarizationabnormality, including any of the following:

-   -   History within 6 months prior to starting study treatment of        myocardial infarction (MI), angina pectoris, coronary artery        bypass graft (CABG).    -   Clinically significant cardiac arrhythmias (e.g., ventricular        tachycardia), complete left bundle branch block, high-grade AV        block (e.g., bifascicular block, Mobitz type II and third degree        AV block).    -   QTcF at screening ≥450 ms (male patients), ≥460 ms (female        patients).    -   Long QT syndrome, family history of idiopathic sudden death or        congenital long QT syndrome, or any of the following:        -   Risk factors for Torsades de Pointes (TdP) including            uncorrected hypokalemia or hypomagnesemia, history of            cardiac failure, or history of clinically            significant/symptomatic bradycardia.        -   Concomitant medication(s) with a known risk to prolong the            QT interval and/or known to cause Torsades de Pointes that            cannot be discontinued or replaced 7 days prior to starting            study drug by safe alternative medication.        -   Inability to determine the QTcF interval.

Preferably, the patient does not have severe and/or uncontrolledconcurrent medical disease that in the opinion of the investigator couldcause unacceptable safety risks or compromise compliance with theprotocol (e.g., uncontrolled diabetes, active or uncontrolled infection,pulmonary hypertension).

As used herein, IL-1β inhibitors include but not be limited to,canakinumab or a functional fragment thereof, gevokizumab or afunctional fragment thereof, Anakinra, diacerein, Rilonacept, IL-1Affibody (SOBI 006, Z-FC (Swedish Orphan Biovitrum/Affibody)) andLutikizumab (ABT-981) (Abbott), CDP-484 (Celltech), LY-2189102 (Lilly).

In one embodiment of any use or method of the invention, said IL-1βbinding antibody is canakinumab. Canakinumab (ACZ885) is ahigh-affinity, fully human monoclonal antibody of the IgG1/k tointerleukin-1β, developed for the treatment of IL-1β driven inflammatorydiseases. It is designed to bind to human IL-1β and thus blocks theinteraction of this cytokine with its receptors. Canakinumab isdisclosed in WO02/16436 which is hereby incorporated by reference in itsentirety.

In other embodiments of any use or method of the invention, said IL-1βbinding antibody is gevokizumab. Gevokizumab (XOMA-052) is ahigh-affinity, humanized monoclonal antibody of the IgG2 isotype tointerleukin-1β, developed for the treatment of IL-1β driven inflammatorydiseases. Gevokizumab modulates IL-1β binding to its signaling receptor.Gevokizumab is disclosed in WO2007/002261 which is hereby incorporatedby reference in its entirety.

In one embodiment said IL-1β binding antibody is LY-2189102, which is ahumanised interleukin-1 beta (IL-1β) monoclonal antibody.

In one embodiment said IL-1β binding antibody or a functional fragmentthereof is CDP-484 (Celltech), which is an antibody fragment blockingIL-1β.

In one embodiment said IL-1β binding antibody or a functional fragmentthereof is IL-1 Affibody (SOBI 006, Z-FC (Swedish OrphanBiovitrum/Affibody)).

An antibody, as used herein, refers to an antibody having the naturalbiological form of an antibody. Such an antibody is a glycoprotein andconsists of four polypeptides—two identical heavy chains and twoidentical light chains, joined to form a “Y”-shaped molecule. Each heavychain is comprised of a heavy chain variable region (VH) and a heavychain constant region. The heavy chain constant region is comprised ofthree or four constant domains (CH1, CH2, CH3, and CH4, depending on theantibody class or isotype). Each light chain is comprised of a lightchain variable region (VL) and a light chain constant region, which hasone domain, CL. Papain, a proteolytic enzyme, splits the “Y” shape intothree separate molecules, two so called “Fab” fragments (Fab=fragmentantigen binding), and one so called “Fc” fragment (Fc=fragmentcrystallizable). A Fab fragment consists of the entire light chain andpart of the heavy chain. The VL and VH regions are located at the tipsof the “Y”-shaped antibody molecule. The VL and VH each have threecomplementarity-determining regions (CDRs).

By “IL-1β binding antibody” is meant any antibody capable of binding tothe IL-1β specifically and consequently inhibiting or modulating thebinding of IL-1β to its receptor and further consequently inhibitingIL-1β function. Preferably an IL-1β binding antibody does not bind toIL-1α.

Preferably an IL-1β binding antibody includes:

(1) An antibody comprising three VL CDRs having the amino acid sequencesRASQSIGSSLH (SEQ ID NO: 1), ASQSFS (SEQ ID NO: 2), and HQSSSLP (SEQ IDNO: 3) and three VH CDRs having the amino acid sequences VYGMN (SEQ IDNO: 5), IIWYDGDNQYYADSVKG (SEQ ID NO: 6), and DLRTGP (SEQ ID NO: 7);

(2) An antibody comprising three VL CDRs having the amino acid sequencesRASQDISNYLS (SEQ ID NO: 9), YTSKLHS (SEQ ID NO: 10), and LQGKMLPWT (SEQID NO: 11), and three VH CDRs having the amino acid sequences TSGMGVG(SEQ ID NO: 13), HIWWDGDESYNPSLK (SEQ ID NO: 14), and NRYDPPWFVD (SEQ IDNO: 15); and

(3) An antibody comprising the six CDRs as described in either (1) or(2), wherein one or more of the CDR sequences, preferably at most two ofthe CDRs, preferably only one of the CDRs, differ by one amino acid fromthe corresponding sequences described in either (1) or (2),respectively.

Preferably an IL-1β binding antibody includes:

(1) An antibody comprising three VL CDRs having the amino acid sequencesRASQSIGSSLH (SEQ ID NO: 1), ASQSFS (SEQ ID NO: 2), and HQSSSLP (SEQ IDNO: 3) and comprising the VH having the amino acid sequence specified inSEQ ID NO: 8;

(2) An antibody comprising the VL having the amino acid sequencespecified in SEQ ID NO: 4 and comprising three VH CDRs having the aminoacid sequences VYGMN (SEQ ID NO: 5), IIWYDGDNQYYADSVKG (SEQ ID NO: 6),and DLRTGP (SEQ ID NO: 7);

(3) An antibody comprising three VL CDRs having the amino acid sequencesRASQDISNYLS (SEQ ID NO: 9), YTSKLHS (SEQ ID NO: 10), and LQGKMLPWT (SEQID NO: 11), and comprising the VH having the amino acid sequencesspecified in SEQ ID NO: 16;

(4) An antibody comprising the VL having the amino acid specified in SEQID NO: 12, and comprising three VH CDRs having the amino acid sequencesTSGMGVG (SEQ ID NO: 13), HIWWDGDESYNPSLK (SEQ ID NO: 14), and NRYDPPWFVD(SEQ ID NO: 15);

(5) An antibody comprising three VL CDRs and the VH sequence asdescribed in either (1) or (3), wherein one or more of the VL CDRsequences, preferably at most two of the CDRs, preferably only one ofthe CDRs, differ by one amino acid from the corresponding sequencesdescribed in (1) or (3), respectively, and wherein the VH sequence is atleast 90% identical to the corresponding sequence described in (1) or(3), respectively; and

(6) An antibody comprising the VL sequence and three VH CDRs asdescribed in either (2) or (4), wherein the VL sequence is at least 90%identical to the corresponding sequence described in (2) or (4),respectively, and wherein one or more of the VH CDR sequences,preferably at most two of the CDRs, preferably only one of the CDRs,differ by one amino acid from the corresponding sequences described in(2) or (4), respectively.

Preferably an IL-1β binding antibody includes:

(1) An antibody comprising the VL having the amino acid sequencespecified in SEQ ID NO: 4 and comprising the VH having the amino acidsequence specified in SEQ ID NO: 8;

(2) An antibody comprising the VL having the amino acid specified in SEQID NO: 12, and comprising the VH having the amino acid sequencesspecified in SEQ ID NO: 16; and

(3) An antibody described in either (1) or (2), wherein the constantregion of the heavy chain, the constant region of the light chain orboth has been changed to a different isotype as compared to canakinumabor gevokizumab.

Preferably an IL-1β binding antibody includes:

(1) Canakinumab (SEQ ID NO:17 and 18); and

(2) Gevokizumab (SEQ ID NO:19 and 20).

An IL-1β binding antibody as defined above has substantially identicalor identical CDR sequences as those of canakinumab or gevokizumab. Itthus binds to the same epitope on IL-1β and has similar binding affinityas canakinumab or gevokizumab. The clinical relevant doses and dosingregimens that have been established for canakinumab or gevokizumab astherapeutically efficacious in the treatment of cancer, especiallycancer having at least partial inflammatory basis, would be applicableto other IL-1β binding antibodies.

Additionally or alternatively, an IL-1β antibody refers to an antibodythat is capable of binding to IL-1β specifically with affinity in thesimilar range as canakinumab or gevokizumab. The Kd for canakinumab inWO2007/050607 is referenced with 30.5 pM, whereas the Kd for gevokizumabis 0.3 pM. Thus affinity in the similar range refers to between about0.05 pM to about 300 pM, preferably about 0.1 pM to about 100 pM.Although both binding to IL-1β, canakinumab directly inhibits thebinding to IL-1 receptor, whereas gevokizumab is an allostericinhibitor. It does not prevent IL-1β from binding to the receptor butprevent receptor activation. Preferably an IL-1β antibody has thebinding affinity in the similar range as canakinumab, preferably in therange of about 1 pM to about 300 pM, preferably in the range of about 10pM to about 100 pM, wherein preferably said antibody directly inhibitsbinding. Preferably an IL-1β antibody has the binding affinity in thesimilar range as gevokizumab, preferably in the range of about 0.05 pMto about 3 pM, preferably in the range of about 0.1 pM to about 1 pM,wherein preferably said antibody is an allosteric inhibitor.

As used herein, the term “functional fragment” of an antibody as usedherein, refers to portions or fragments of an antibody that retain theability to specifically bind to an antigen (e.g., IL-1β). Examples ofbinding fragments encompassed within the term “functional fragment” ofan antibody include single chain Fv (scFv), a Fab fragment, a monovalentfragment consisting of the V_(L), V_(H), CL and CH1 domains; a F(ab)2fragment, a bivalent fragment comprising two Fab fragments linked by adisulfide bridge at the hinge region; a Fd fragment consisting of theV_(H) and CH1 domains; a Fv fragment consisting of the V_(L) and V_(H)domains of a single arm of an antibody; a dAb fragment (Ward et al.,1989), which consists of a V_(H) domain; and an isolated complementaritydetermining region (CDR); and one or more CDRs arranged on peptidescaffolds that can be smaller, larger, or fold differently to a typicalantibody.

The term “functional fragment” might also refer to one of the following:

-   -   bispecific single chain Fv dimers (PCT/US92/09965)    -   “diabodies” or “triabodies”, multivalent or multispecific        fragments constructed by gene fusion (Tomlinson I & Hollinger        P (2000) Methods Enzymol. 326: 461-79; W094113804; Holliger P et        al., (1993) Proc. Natl. Acad. Sci. USA, 90: 6444-48)    -   scFv genetically fused to the same or a different antibody        (Coloma M J & Morrison S L (1997) Nature Biotechnology, 15(2):        159-163)    -   scFv, diabody or domain antibody fused to an Fc region    -   scFv fused to the same or a different antibody    -   Fv, scFv or diabody molecules may be stabilized by the        incorporation of disulphide bridges linking the VH and VL        domains (Reiter, Y. et al, (1996) Nature Biotech, 14,        1239-1245).    -   Minibodies comprising a scFv joined to a CH3 domain may also be        made (Hu, S. et al, (1996) Cancer Res., 56, 3055-3061).    -   Other examples of binding fragments are Fab′, which differs from        Fab fragments by the addition of a few residues at the carboxyl        terminus of the heavy chain CH1 domain, including one or more        cysteines from the antibody hinge region, and Fab′-SH, which is        a Fab′ fragment in which the cysteine residue(s) of the constant        domains bear a free thiol group

Typically and preferably an functional fragment of an IL-1β bindingantibody is a portion or a fragment of an “IL-1β binding antibody” asdefined above.

Dosing Regimen of the Present Invention

If an IL-1β inhibitor, such as an IL-1β antibody or a functionalfragment thereof, is administered in a dose range that can effectivelyreduce hsCRP level in a patient with cancer having at least partialinflammatory basis, treatment effect of said cancer can possibly beachieved. Dose range, of a particular IL-1β inhibitor, preferably IL-1βantibody or a functional fragment thereof, that can effectively reducehsCRP level is known or can be tested in a clinical setting.

Thus in one embodiment, the present invention comprises administeringthe IL-1β binding antibody or a functional fragment thereof to a patientwith cancer that has at least a partial inflammatory basis in the rangeof about 20 mg to about 400 mg per treatment, preferably in the range ofabout 30 mg to about 400 mg per treatment, preferably in the range ofabout 30 mg to about 200 mg per treatment, preferably in the range ofabout 60 mg to about 200 mg per treatment. In one embodiment the patientwith a cancer that has at least a partial inflammatory basis, includingCML, receives each treatment about every two weeks, about every threeweeks, about every four weeks (monthly), about every 6 weeks, aboutbimonthly (every 2 months), about every nine weeks or about quarterly(every 3 months). In one embodiment the patient receives each treatmentabout every 3 weeks. In one embodiment the patient receives eachtreatment about every 4 weeks. The term “per treatment”, as used in thisapplication and particularly in this context, should be understood asthe total amount of drug received per hospital visit or perself-administration or per administration helped by a health care giver.Normally and preferably the total amount of drug received per treatmentis administered to a patient within about one day, preferably withinabout half a day, preferably within about 4 hours, preferably withinabout 2 hours. In one preferred embodiment the term “per treatment” isunderstood as the drug is administered with one injection, preferably inone dosage.

In practice sometimes the time interval cannot be strictly kept due tothe limitation of the availability of doctor, patient or thedrug/facility. Thus the time interval can slightly vary, normallybetween about 5 days, about 4 days, about 3 days, about 2 days orpreferably about 1 day.

Sometimes it is desirable to quickly reduce inflammation. IL-1βauto-induction has been shown in human mononuclear blood, human vascularendothelial, and vascular smooth muscle cells in vitro and in rabbits invivo where IL-1 has been shown to induce its own gene expression andcirculating IL-1β level (Dinarello et al. 1987, Warner et al. 1987a, andWarner et al. 1987b).

This induction period over 2 weeks by administration of a first dosefollowed by a second dose two weeks after administration of the firstdose is to assure that auto-induction of IL-1β pathway is adequatelyinhibited at initiation of treatment. The complete suppression of IL-1βrelated gene expression achieved with this early high doseadministration, coupled with the continuous canakinumab treatment effectwhich has been proven to last the entire quarterly dosing period used inCANTOS, is to minimize the potential for IL-1β rebound. In addition,data in the setting of acute inflammation suggests that higher initialdoses of canakinumab that can be achieved through induction are safe andprovide an opportunity to ameliorate concern regarding potentialauto-induction of IL-1β and to achieve greater early suppression ofIL-1β related gene expression.

Thus in one embodiment, the present invention, while keeping the abovedescribed dosing schedules, especially envisages the secondadministration of DRUG of the invention is about one week later or atmost about two weeks, preferably about two weeks apart from the firstadministration. Then the third and the further administration willfollowing the schedule of about every 2 weeks, about every 3 weeks,about every 4 weeks (monthly), about every 6 weeks, about bimonthly(every 2 months), about every 9 weeks or about quarterly (every 3months).

In one embodiment, the IL-1β binding antibody is canakinumab, whereincanakinumab is administered to a patient with cancer that has at least apartial inflammatory basis, e.g., CML, in the range of about 100 mg toabout 400 mg, preferably about 200 mg per treatment. In one embodimentthe patient with cancer that has at least a partial inflammatory basis,e.g., CML, receives each treatment about every 2 weeks, about every 3weeks, about every 4 weeks (monthly), about every 6 weeks, aboutbimonthly (every 2 months), about every 9 weeks or about quarterly(every 3 months). In one embodiment the patient with cancer that has atleast a partial inflammatory basis, e.g., CML, receives canakinumababout monthly or about every three weeks. In one embodiment thepreferred dose of canakinumab for patient with cancer that has at leasta partial inflammatory basis, e.g., CML, is about 200 mg about every 3weeks. In one embodiment the preferred dose of canakinumab is about 200mg about monthly. When a safety concern raises, the dose can bedown-titrated, preferably by increasing the dosing interval, preferablyby doubling or tripling the dosing interval. For example about 200 mgmonthly or about every 3 weeks regimen can be changed to about every 2month or about every 6 weeks respectively or about every 3 month orabout every 9 weeks respectively. In an alternative embodiment thepatient with cancer that has at least a partial inflammatory basis,e.g., CML, receives canakinumab at a dose of about 200 mg about everytwo month or about every 6 weeks in the down-titration phase or in themaintenance phase independent from any safety issue or throughout thetreatment phase. In an alternative embodiment the patient with CMLreceives canakinumab at a dose of about 200 mg about every 3 month orabout every 9 weeks in the down-titration phase or in the maintenancephase independent from any safety issue or throughout the treatmentphase. In an alternative embodiment the patient receives canakinumab ata dose of about 250 mg. In an alternative embodiment the patientreceives canakinumab at a dose of about 250 mg about every 4 weeks.

Suitable the above dose and dosing apply to the use of a functionalfragment of canakinumab according to the present invention.

Canakinumab or a functional fragment thereof can be administeredintravenously or subcutaneously, preferably subcutaneously.

The dosing regimens disclosed herein is applicable in each and everycanakinumab related embodiments disclosed in this application, includingbut not limited to monotherapy or in combination with one or morechemotherapeutic agent, used in adjuvant setting or in the first line,2^(nd) line or 3^(rd) line treatment.

In one embodiment, the present invention comprises administeringgevokizumab to a patient with cancer that has at least a partialinflammatory basis, e.g., CML, in the range of about 20 mg to about 240mg per treatment, preferably in the range of about 30 mg to about 180mg, preferably in the range of about 30 mg to about 120 mg, preferablyabout 30 mg to about 60 mg, preferably about 60 mg to about 120 mg pertreatment. In one embodiment the patient receives about 30 mg to about120 mg per treatment. In one embodiment the patient receives about 30 mgto about 60 mg per treatment. In one embodiment the patient receivesabout 30 mg, about 60 mg, about 90 mg, about 120 mg or about 180 mg pertreatment. In one embodiment the patient with cancer that has at least apartial inflammatory basis, e.g., CML, receives each treatment aboutevery 2 weeks, about every 3 weeks, about monthly (every 4 weeks), aboutevery 6 weeks, about bimonthly (every 2 months), about every 9 weeks orabout quarterly (every 3 months). In one embodiment the patient receiveseach treatment about every 3 weeks. In one embodiment the patientreceives each treatment about every 4 weeks.

When a safety concern raises, the dose can be down-titrated, preferablyby increasing the dosing interval, preferably by doubling or triplingthe dosing interval. For example about 60 mg about monthly or aboutevery 3 weeks regimen can be doubled to about every 2 month or aboutevery 6 weeks respectively or tripled to about every 3 month or aboutevery 9 weeks respectively. In an alternative embodiment the patientwith cancer that has at least a partial inflammatory basis, e.g., CML,receives gevokizumab at a dose of about 30 mg to about 120 mg aboutevery 2 month or about every 6 weeks in the down-titration phase or inthe maintenance phase independent from any safety issue or throughoutthe treatment phase. In an alternative embodiment the patient withbreast cancer receives gevokizumab at a dose of about 30 mg to about 120mg about every 3 month or about every 9 weeks in the down-titrationphase or in the maintenance phase independent from any safety issue orthroughout the treatment phase.

Suitably the above dose and dosing apply to the use of a functionalfragment of gevokizumab according to the present invention.

Gevokizumab or a functional fragment thereof can be administeredintravenously or subcutaneously, preferably intravenously.

The dosing regimens disclosed herein is applicable in each and everygevokizumab related embodiments disclosed in this application, includingbut not limited to monotherapy or in combination with one or morechemotherapeutic agent, used in adjuvant setting or in the first line,2^(nd) line or 3^(rd) line treatment.

Biomarkers

In one aspect, the present invention provides the use of an IL-1βbinding antibody or a functional fragment thereof, suitably canakinumabor a functional fragment thereof, gevokizumab or a functional fragmentthereof, in the treatment and/or prevention of cancer, e.g., cancerhaving at least a partial inflammatory basis, in a patient who has ahigher than normal level of C-reactive protein (hsCRP). In one furtherembodiment, this patient is a smoker. In one further embodiment, thepatient is a current smoker. Typically cancers that have at least apartial inflammatory basis that possibly have patients exhibiting higherthan normal hsCRP levels include, but are not limited to, CML.

As used herein, “C-reactive protein” and “CRP” refers to serum or plasmaC-reactive protein, which is typically used as an indicator of the acutephase response to inflammation. Nonetheless, CRP level may becomeelevated in chronic illnesses such as cancer. The level of CRP in serumor plasma may be given in any concentration, e.g., mg/dl, mg/L, nmol/L.Levels of CRP may be measured by a variety of well-known methods, e.g.,radial immunodiffusion, electroimmunoassay, immunoturbidimetry (e.g.,particle (e.g., latex)-enhanced turbidimetric immunoassay), ELISA,turbidimetric methods, fluorescence polarization immunoassay, and lasernephelometry. Testing for CRP may employ a standard CRP test or a highsensitivity CRP (hsCRP) test (i.e., a high sensitivity test that iscapable of measuring lower levels of CRP in a sample, e.g., usingimmunoassay or laser nephelometry). Kits for detecting levels of CRP maybe purchased from various companies, e.g., Calbiotech, Inc, CaymanChemical, Roche Diagnostics Corporation, Abazyme, DADE Behring, AbnovaCorporation, Aniara Corporation, Bio-Quant Inc., Siemens HealthcareDiagnostics, Abbott Laboratories etc.

As used herein, the term “hsCRP” refers to the level of CRP in the blood(serum or plasma) as measured by high sensitivity CRP testing. Forexample, Tina-quant C-reactive protein (latex) high sensitivity assay(Roche Diagnostics Corporation) may be used for quantification of thehsCRP level of a subject. Such latex-enhanced turbidimetric immunoassaymay be analysed on the Cobas® platform (Roche Diagnostics Corporation)or Roche/Hitachi (e.g., Modular P) analyzer. In the CANTOS trial thehsCRP level was measured by Tina-quant C-reactive protein (latex) highsensitivity assay (Roche Diagnostics Corporation) on the Roche/HitachiModular P analyzer, which can be used typically and preferably as themethod in measuring hsCRP level. Alternatively the hsCRP level can bemeasured by another method, for example by another approved companiondiagnostic kit, the value of which can be calibrated against the valuemeasured by the Tina-quant method.

Each local laboratory employ a cut-off value for abnormal (high) CRP orhsCRP based on that laboratory's rule for calculating normal maximumCRP, i.e. based on that laboratory's reference standard. A physiciangenerally orders a CRP test from a local laboratory, and the locallaboratory determines CRP or hsCRP value and reports normal or abnormal(low or high) CRP using the rule that particular laboratory employs tocalculate normal CRP, namely based on its reference standard. Thuswhether a patient has a higher than normal level of C-reactive protein(hsCRP) can be determined by the local laboratory where the test isconducted.

It is plausible that an IL-1β antibody or a fragment thereof, such ascanakinumab or gevokizumab, is effective in treating and/or preventingother cancer having at least partially inflammatory basis in a patient,especially when said patient has higher than normal level of hsCRP. Likecanakinumab, gevokizumab binds to IL-1β specifically. Unlike canakinumabdirectly inhibiting the binding of IL-1β to its receptor, gevokizumab isan allosteric inhibitor. It does not inhibit IL-1β from binding to itsreceptor but prevent the receptor from being activated by IL-1β. Likecanakinumab, gevokizumab was tested in a few inflammation basedindications and has been shown to effectively reduce inflammation asindicated, for example, by the reduction of hsCRP level in thosepatients. Furthermore from the available IC50 value, gevokizumab seemsto be a more potent IL-1β inhibitor than canakinumab.

Furthermore, the present invention provides effective dosing ranges,within which the hsCRP level can be reduced to a certain threshold,below which more patients with cancer having at least partiallyinflammatory basis can become responder or below which the same patientcan benefit more from the great therapeutic effect of the DRUG of theinvention with negligible or tolerable side effects.

In one aspect, the present invention provides high sensitivityC-reactive protein (hsCRP) or CRP for use as a biomarker in thetreatment of cancer, e.g., cancer having at least a partial inflammatorybasis, e.g., CML, with an IL-1β inhibitor, e.g., IL-1β binding antibodyor a functional fragment thereof. The level of hsCRP is possiblyrelevant in determining whether a patient with diagnosed or undiagnosedcancer should be treated with an IL-1β binding antibody or a functionalfragment thereof. In one embodiment patient is eligible for thetreatment if the level of hsCRP is equal to or higher than about 2.5mg/L, or equal to or higher than about 4.5 mg/L, or equal to or higherthan about 7.5 mg/L, or equal to or higher than about 9.5 mg/L, asassessed prior to the administration of the IL-1β binding antibody or afunctional fragment thereof.

In one embodiment, the present invention provides the use of an IL-1βbinding antibody or a functional fragment thereof, suitably canakinumabor gevokizumab, for the treatment and/or prevention of cancer, e.g.,cancer that has at least a partial inflammatory basis in a patient whohas high sensitivity C-reactive protein (hsCRP) level equal to or higherthan about 2 mg/L, equal to or higher than about 3 mg/L, equal to orhigher than about 4 mg/L, equal to or higher than about 5 mg/L, equal toor higher than about 6 mg/L, equal to or higher than about 7 mg/L, equalto or higher than about 8 mg/L, equal to or higher than about 9 mg/L,equal to or higher than about 10 mg/L, equal to or higher than about 12mg/L, equal to or higher than about 15 mg/L, equal to or higher thanabout 20 mg/L or equal to or higher than about 25 mg/L, preferablybefore first administration of said IL-1β binding antibody or functionalfragment thereof. Preferably said patient has a hsCRP level equal to orhigher than about 4 mg/L. Preferably said patient has a hsCRP levelequal to or higher than about 6 mg/L. Preferably said patient has ahsCRP level equal to or higher than about 10 mg/L. Preferably saidpatient has a hsCRP level equal to or higher than about 20 mg/L. In onefurther embodiment, this patient is a smoker. In one further embodiment,this patient is a current smoker.

In one embodiment, the present invention provides the use of an IL-1βbinding antibody or a functional fragment thereof, suitably canakinumabor gevokizumab, for the treatment of cancer, e.g., cancer that has atleast a partial inflammatory basis in a patient who has a highsensitivity C-reactive protein (hsCRP) level equal to or higher about 6mg/L, equal to or higher than about 10 mg/L or equal to or higher thanabout 15 mg/L, preferably before first administration of DRUG of theinvention. In one embodiment said cancer is CRC, RCC, pancreatic cancer,melanoma, HCC or gastric cancer.

In one aspect the present invention provides an IL-1β binding antibodyor a functional fragment thereof for use in the treatment of cancer,e.g., cancer having at least a partial inflammatory basis, e.g., CML, ina patient, wherein the efficacy of the treatment correlates with thereduction of hsCRP in said patient, comparing to prior treatment. In oneembodiment the present invention provides an IL-1β binding antibody or afunctional fragment thereof for use in the treatment of cancer, e.g.,cancer having at least a partial inflammatory basis, e.g., CML, in apatient, wherein the CRP level, more precisely the hsCRP level, of saidpatient has reduced to below about 9 mg/L, preferably to below about 2.4mg/L, preferably to below about 2 mg/L, to below about 1.8 mg/L, about 6months, or preferably about 3 months from the first administration ofsaid IL-1β binding antibody or a functional fragment thereof at a properdose, preferably according to the dosing regimen of the presentinvention.

In one aspect, the present invention provides IL-6 use as a biomarker inthe treatment and/or prevention of cancer, e.g., cancer having at leasta partial inflammatory basis, e.g., CML, with an IL-1β inhibitor, e.g.,IL-1β binding antibody or a functional fragment thereof. The level ofIL-6 is possibly relevant in determining whether a patient withdiagnosed or undiagnosed cancer or is at risk of developing cancershould be treated with an IL-1β binding antibody or a functionalfragment thereof. In one embodiment patient is eligible for thetreatment and/or prevention if the level of IL-6 is equal to or higherthan about 1.9 pg/ml, higher than about 2 pg/ml, higher than about 2.2pg/ml, higher than about 2.5 pg/ml, higher than about 2.7 pg/ml, higherthan about 3 pg/ml, higher than about 3.5 pg/ml, as assessed prior tothe administration of the IL-1β binding antibody or a functionalfragment thereof. Preferably the patient has an IL-6 level equal to orhigher than about 2.5 mg/L.

In one aspect the present invention provides an IL-1β binding antibodyor a functional fragment thereof for use in the treatment and/orprevention of cancer, e.g., cancer having at least a partialinflammatory basis, e.g., CML, in a patient, wherein the efficacy of thetreatment correlates with the reduction of IL-6 in said patient,comparing to prior treatment. In one embodiment the present inventionprovides an IL-1β binding antibody or a functional fragment thereof foruse in the treatment of cancer, e.g., cancer having at least a partialinflammatory basis, e.g., CML, wherein hsCRP level of said patient hasreduced to below about 2.2 pg/ml, preferably to below about 2 pg/ml,preferably to below about 1.9 pg/ml about 6 months, or preferably about3 months from the first administration of said IL-1β binding antibody ora functional fragment thereof at a proper dose, preferably according tothe dosing regimen of the present invention.

In one embodiment, said cancer is CML.

In one aspect the present invention provides an IL-1β binding antibodyor a functional fragment thereof (e.g., canakinumab or gevokizumab) foruse in the treatment of cancers that have at least a partialinflammatory basis, e.g., CML, in a patient, wherein the hsCRP level ofsaid patient has reduced by at least about 15%, at least about 20%, atleast about 30% or at least about 40% about 6 months, or preferablyabout 3 month from the first administration of said IL-1β bindingantibody or a functional fragment thereof at a proper dose, preferablyaccording to the dosing regimen of the present invention, as compared tothe hsCRP level just prior to the first administration of the IL-1βbinding antibody or a functional fragment thereof, canakinumab orgevokizumab). Further preferably the hsCRP level of said patient hasreduced by at least about 15%, at least about 20%, at least about 30%after the first administration of the DRUG of the invention according tothe dose regimen of the present invention.

In one aspect the present invention provides an IL-1β binding antibodyor a functional fragment thereof (e.g., canakinumab or gevokizumab) foruse in the treatment of cancers, e.g., cancers that have at least apartial inflammatory basis, e.g., CML, in a patient, wherein the IL-6level of said patient has reduced by at least about 15%, at least about20%, at least about 30% or at least about 40% about 6 months, orpreferably about 3 months from the first administration of said IL-1βbinding antibody or a functional fragment thereof (e.g., canakinumab orgevokizumab) at a proper dose, preferably according to the dosingregimen of the present invention, as compared to the IL-6 level justprior to the first administration. The term “about” used herein includesa variation of ±10 days from the 3 months or a variation of ±15 daysfrom the 6 months. In one embodiment said cancer is CML.

The reduction of the level of hsCRP and the reduction of the level ofIL-6 can be used separately or in combination to indicate the efficacyof the treatment or as prognostic markers.

Inhibition of Angiogenesis

In one aspect, the present invention provides an IL-1β binding antibodyor a functional fragment thereof, suitably canakinumab or gevokizumab,for use in a patient in need thereof in the treatment of cancer, e.g., acancer having at least partial inflammatory basis, e.g., CML, wherein atherapeutic amount is administered to inhibit angiogenesis in saidpatient. Without wishing to be bound by theory, it is hypothesized thatthe inhibition of IL-1β pathway can lead to inhibition or reduction ofangiogenesis, which is a key event for tumor growth and for tumormetastasis. In clinical settings the inhibition or reduction ofangiogenesis can be measured by tumor shrinkage, no tumor growth (stabledisease), prevention of metastasis or delay of metastasis.

All the disclosed uses throughout this application, including but arenot limited to, doses and dosing regimens, combinations, routes ofadministration and biomarkers can be applied to the aspect of inhibitionor reduction of angiogenesis. In one embodiment canakinumab orgevokizumab are used in combination of one or more anti-cancertherapeutic agents. In one embodiment the one or more chemotherapeuticagents is an anti-Wnt inhibitor, preferably Vantictumab. In oneembodiment the one or more therapeutic agents is a VEGF inhibitor,preferably sunitinib, sorafenib, axitinib, pazopanib, bevacizumab orRamucirumab.

Inhibition of Metastasis

In one embodiment, said IL-1β binding antibody is canakinumab or afunctional fragment thereof. In one preferred embodiment, canakinumab isadministered at a dose of about 200 mg. In one preferred embodiment,canakinumab is administered at a dose of about 200 mg about every 3weeks or about monthly. In one preferred embodiment, canakinumab isadministered at a dose of about 200 mg about every 3 weeks or aboutmonthly subcutaneously. In another embodiment, said IL-1β bindingantibody is gevokizumab or a functional fragment thereof. In onepreferred embodiment, the proper dose of the first administration ofgevokizumab is about 180 mg. In one preferred embodiment, gevokizumab isadministered at a dose of about 60 mg to about 90 mg. In one preferredembodiment, gevokizumab is administered at a dose of about 60 mg toabout 90 mg about every 3 weeks or about monthly. In one preferredembodiment, gevokizumab is administered at a dose of about 120 mg aboutevery 3 weeks or about every 4 weeks (monthly). In one preferredembodiment, gevokizumab is administered intravenously. In one preferredembodiment, gevokizumab is administered at a dose of about 120 mg aboutevery 3 weeks or about every 4 weeks (monthly) intravenously. In onepreferred embodiment, gevokizumab is administered at a dose of about 90mg about every 3 weeks or about every 4 weeks (monthly) intravenously.

Without wishing to be bound by the theory, it is hypothesized thatchronic inflammation, either local or systematic, especially localinflammation, creates an immunosuppressive microenvironment thatpromotes tumor growth and dissemination. IL-1β binding antibody or afunctional fragment thereof reduces chronic inflammation, especiallyIL-1β mediated chronic inflammation, and thereby prevents or delays theoccurrence of cancer in a subject who has otherwise local or systematicchronic inflammation.

One way of determining local or systematic chronic inflammation isthrough measuring the level of C-reactive protein (hsCRP).

In one preferred embodiment, canakinumab is administered at a dose ofabout 100 mg to about 400 mg, preferably about 200 mg about monthly,about every other month or about quarterly, preferably subcutaneously orpreferably about 100 mg about monthly, about every other month or aboutquarterly, preferably subcutaneously. In another embodiment, said IL-1βbinding antibody is gevokizumab or a functional fragment thereof. In onepreferred embodiment, gevokizumab is administered at a dose of about 15mg to about 120 mg. In one preferred embodiment, gevokizumab isadministered about monthly, about every other month or about quarterly.In one preferred embodiment, gevokizumab is administered at a dose ofabout 15 mg about monthly, about every other month or about quarterly.In one preferred embodiment, gevokizumab is administered at a dose ofabout 30 mg about monthly, about every other month or about quarterly.In one embodiments gevokizumab is administered subcutaneously. In oneembodiments gevokizumab is administered intravenously.

Adjuvant Treatment

In one aspect, the present invention provides an IL-1β binding antibodyor a functional fragment thereof, suitably canakinumab or a functionalfragment thereof, or gevokizumab or a functional fragment thereof, foruse in the prevention of recurrence or relapse of cancer, e.g., cancerhaving at least a partial inflammatory basis, which has been surgicallyremoved (resected “adjuvant chemotherapy”). In one embodiment, cancerhaving at least a partial inflammatory basis is not lung cancer,especially not NSCLC.

Without wishing to be bound by the theory, after tumor is surgicallyremoved, it is possible that the inflammation is greatly reduced due tosurgery. The IL-1β or the hsCRP level is no longer higher than normal.It is however reasonable to expect that the DRUG of the invention canprevent or delay the recurrence or relapse of cancer by keepinginflammation under control and thereby preventing the re-formation ofIL-1β mediated immune suppressive tumor microenvironment that promotestumor growth and metastasis. Furthermore after a tumor has beensurgically removed, the patient's immune system can regain itssurveillance function in eliminating remaining tumor loci/cells. Byreducing inflammation, IL-1β binding antibody or a functional fragmentthereof helps maintaining or improving the surveillance function of theimmune system and thereby prevents or delays tumor recurrence or relapseof cancer.

In one embodiment said patient has completed post-surgery standard ofcare adjuvant (other than the treatment of DRUG of the invention)treatment, typically as standard adjuvant chemotherapy, and/or standardadjuvant radiotherapy treatment. The term “post-surgery standard of careadjuvant treatment” includes standard of care small moleculechemotherapeutic agents as post-surgery adjuvant treatment. In oneembodiment the post-surgery SoC adjuvant treatment is cisplatin-baseddoublet chemotherapy. In one further embodiment the post-surgery SoCadjuvant treatment includes 4 cycles of ducisplatin-based doubletchemotherapy treatment.

In one embodiment the patient has just completed the post-surgerystandard of care adjuvant (other than the treatment of DRUG of theinvention) treatment. The term “just” means the interval between thelast administering of the SoC adjuvant drug or the last SoC radiotherapytreatment and the first administration of the DRUG of the invention isnot longer than about 2 months, preferably not longer than 1 month. Inone embodiment the patient receives the first dose of DRUG of theinvention at least about 3 months or at least about 6 months from hislast radiotherapy or his last administering of the SoC adjuvant drug.

In one embodiment the patient only receives DRUG of the invention afterthe patient has completed his SoC of adjuvant treatment. In one furtherembodiment the patient receives DRUG of the invention for at least about6 months, preferably for at least about 12 months, preferably for about12 months. Due to the good safety profile of the DRUG of the invention,especially canakinumab or gevokizumab, the patient receives DRUG of theinvention for at least about 24 months, preferably till the recurrenceor relapse of cancer.

In one embodiment DRUG of the invention is added on top of thepost-surgery standard of care adjuvant treatment, preferably DRUG of theinvention is administered at the beginning of the patient's post-surgerySoC adjuvant treatment. In one further embodiment DRUG of the inventionis continued, preferably as monotherapy, after patient has completed hispost-surgery SoC adjuvant treatment.

In one embodiment the present invention provides an IL-1β bindingantibody or a functional fragment thereof, suitably canakinumab or afunctional fragment thereof, or gevokizumab or a functional fragmentthereof, for use in the prevention of recurrence or relapse of cancer,e.g., cancer having at least a partial inflammatory basis, which hasbeen surgically removed (resected “adjuvant chemotherapy”), wherein thepatient does not receive the post-surgery SoC adjuvant treatment orcould not have completed the post-surgery SoC adjuvant treatment,possibly due to intolerance. In this case, DRUG of the invention is thesole post-surgery adjuvant treatment.

In the adjuvant settings, DRUG of the invention is administeredaccording to the dosing regimen of the present invention. When used asmonotherapy, the dosing interval can be flexible. For example, DRUG ofthe invention can be administered in the loading phase and in themaintenance phase, wherein the average dose is higher in the loadingphase than the that in the maintenance phase. For example DRUG of theinvention can be administered about every 3 weeks or about monthlypost-surgery in the loading phase. The dose interval can be doubled ortripled in the maintenance phase. In one embodiment the loading phase isat least about 6 months, preferably at least about 12 months, preferablyabout 12 months. In one embodiment the maintenance dose is at leastabout 12 months, preferably till the recurrence or relapse of thecancer.

In one embodiment, the present invention provides an IL-1β bindingantibody or a functional fragment thereof, suitably canakinumab or afunctional fragment thereof, or gevokizumab or a functional fragmentthereof, for use in the prevention of recurrence or relapse of cancer,e.g., cancer having at least a partial inflammatory basis, which hasbeen surgically removed (resected “adjuvant chemotherapy”), wherein thedisease free survival (DFS) is at least about 6 months or at least about9 months, or at least about 12 months longer than patients did notreceive DRUG of the invention as adjuvant treatment. DFS is defined asthe time from the date of randomization to the date of detection offirst disease recurrence. In one embodiment patient is followed up everyabout 12 weeks after the completion of the adjuvant treatment of thepresent invention. In one embodiment detection of first diseaserecurrence will be done by clinical evaluation that includes physicalexamination, and radiological tumor measurements as determined by theinvestigator.

In one embodiment the overall survival (OS, defined as the time from thedate of randomization to the date of death due to any cause) is at leastabout 6 months, preferably at least about 12 months longer than patientsdid not receive DRUG of the invention as adjuvant treatment.

First Line Treatment

In one embodiment, the present invention provides an IL-1β antibody or afunctional fragment thereof, suitably canakinumab or gevokizumab, foruse as the first line treatment of cancer, e.g., cancer having at leasta partial inflammatory basis e.g., CML. The term “first line treatment”means said patient is given the IL-1β antibody or a functional fragmentthereof before the patient develops resistance to the initial treatmentwith one or more other therapeutic agents. Preferably one or more othertherapeutic agents is a platinum-based mono or combination therapy, atargeted therapy, such a tyrosine inhibitor therapy, a checkpointinhibitor therapy or any combination thereof. As first line treatment,the IL-1β antibody or a functional fragment thereof, such as canakinumabor gevokizumab, can be administered to the patient as monotherapy orpreferably in combination with one or more therapeutic agents, such as acheck point inhibitor, particularly a PD-1 or PD-L1 inhibitor,preferably pembrolizumab, with or without one or more small moleculechemotherapeutic agent. In one embodiment as first line treatment, theIL-1β antibody or a functional fragment thereof, such as canakinumab orgevokizumab, can be administered to the patient in combination with thestandard of care therapy for that cancer. Preferably canakinumab orgevokizumab is administered as the first line treatment until diseaseprogression.

Second Line Treatment

In one embodiment, the present invention provides an IL-1β antibody or afunctional fragment thereof, suitably canakinumab or gevokizumab, foruse as the second or third line treatment of cancer, e.g., cancer havingat least a partial inflammatory basis, e.g., CML. The term “the secondor third line treatment” means IL-1β antibody or a functional fragmentthereof is administered to a patient with cancer progression on or afterone or more other therapeutic agent treatment, especially diseaseprogression on or after FDA-approved first line therapy for that cancer.Preferably one or more other therapeutic agent is a chemotherapeuticagent, such as platinum-based mono or combination therapy, a targetedtherapy, such a tyrosine inhibitor therapy, a checkpoint inhibitortherapy or any combination thereof.

In one embodiment, the IL-1β binding antibody or a functional fragmentthereof, suitably canakinumab or gevokizumab, is administered to saidpatient with cancer having at least partial inflammatory basis prior tosurgery (neoadjuvant chemotherapy) or post-surgery (adjuvantchemotherapy). In one embodiment, IL-1β binding antibody or functionalfragment thereof is administered to said patient prior to, concomitantlywith or post radiotherapy.

In one embodiment DRUG of the invention, suitably canakinumab orgevokizumab, can be used in 2, 3 or all lines of the treatment of cancerin the same patient. Treatment line typically includes but not limitedto neo-adjuvant treatment, adjuvant treatment, first line treatment,2^(nd) line treatment, 3^(rd) line treatment and further line oftreatment. Patient normally changes lines of treatment after surgery,after disease progression or after developing drug resistance to thecurrent treatment. In one embodiment DRUG of the invention is continuedafter patient develops resistant to the current treatment. In oneembodiment DRUG of the invention is continued to the next line oftreatment. In one embodiment DRUG of the invention is continued afterdisease progression. In one embodiment DRUG of the invention iscontinued until death or until palliative care.

In one embodiment the present invention provides DRUG of the invention,suitably canakinumab or gevokizumab, for use in re-treating cancer in apatient, wherein the patient was treated with the same DRUG of theinvention in the previous treatment. In one embodiment the previoustreatment is the neo-adjuvant treatment. In one embodiment the previoustreatment is the adjuvant treatment. In one embodiment the previoustreatment is the first line treatment. In one embodiment the previoustreatment is the second line treatment.

Combination

In one aspect, the present invention provides an IL-1β binding antibodyor a functional fragment thereof, suitably canakinumab or a functionalfragment thereof, or gevokizumab or a functional fragment thereof, foruse in a patient in need thereof in the treatment of a cancer,particularly cancer having at least partial inflammatory basis, e.g.,CML, wherein said IL-1β binding antibody or a functional fragmentthereof is administered in combination with one or more therapeuticagent, e.g., chemotherapeutic agents.

Without wishing to be bound by theory, it is believed that typicalcancer development requires two steps. Firstly, gene alteration resultsin cell growth and proliferation no longer subject to regulation.Secondly, the abnormal tumor cells evade surveillance of the immunesystem. Inflammation plays an important role in the second step.Therefore, control of inflammation can stop cancer development at theearly or earlier stage. Thus it is expected that blocking the IL-1βpathway to reduce inflammation would have a general benefit,particularly improvement of the treatment efficacy on top of thestandard of care, which is normally mainly to directly inhibit thegrowth and proliferation of the malignant cells. In one embodiment, theone or more therapeutic agent, e.g., chemotherapeutic agents is thestandard of care agents of said cancer, particularly cancer having atleast partial inflammatory basis.

Check point inhibitors de-supress the immune system through a mechanismdifferent from IL-1β inhibitors. Thus the addition of IL-1β inhibitors,particularly IL-1β binding antibodies or a functional fragment thereofto the standard Check point inhibitors therapy will further active theimmune response, particularly at the tumor microenvironment.

In one embodiment, the one or more therapeutic agents is nivolumab.

In one embodiment, the one or more therapeutic agents is pembrolizumab.

In one embodiment, the one or more therapeutic agent is nivolumab andipilimumab.

In one embodiment, the one or more chemotherapeutic agents iscabozantinib, or a pharmaceutically acceptable salt thereof.

In one embodiment the or more therapeutic agent is Atezolizumab plusbevacizumab.

In one embodiment, the one or more chemotherapeutic agents isbevacizumab.

In one embodiment, the one or more chemotherapeutic agents is FOLFIRI,FOLFOX or XELOX.

In one embodiment the one or more chemotherapeutic agent is FOLFIRI plusbevacizumab or FOLFOX plus bevacizumab.

In one embodiment the one or more chemotherapeutic agent isplatinum-based doublet chemotherapy (PT-DC).

Therapeutic agents are cytotoxic and/or cytostatic drugs (drugs thatkill malignant cells, or inhibit their proliferation, respectively) aswell as check point inhibitors. Chemotherapeutic agents can be, forexample, small molecule agents, biologics agents (e.g., antibodies, celland gene therapies, cancer vaccines), hormones or other natural orsynthetic peptide or polypeptides. Commonly known chemotherapeutic agentincludes, but is not limited to, platinum agents (e.g., cisplatin,carboplatin, oxaliplatin, nedaplatin, triplatin, lipoplatin,satraplatin, picoplatin), antimetabolites (e.g., methotrexate,5-Fluorouracil, gemcitabine, pemetrexed, edatrexate), mitotic inhibitors(e.g., paclitaxel, albumin-bound paclitaxel, docetaxel, taxotere,docecad), alkylating agents (e.g., cyclophosphamide, mechlorethaminehydrochloride, ifosfamide, melphalan, thiotepa), vinca alkaloids (e.g.,vinblastine, vincristine, vindesine, vinorelbine), topoisomeraseinhibitors (e.g., etoposide, teniposide, topotecan, irinotecan,camptothecin, doxorubicin), antitumor antibiotics (e.g., mitomycin C)and/or hormone-modulating agents (e.g., anastrozole, tamoxifen).Examples of anti-cancer agents used for chemotherapy includeCyclophosphamide (Cytoxan®), Methotrexate, 5-Fluorouracil (5-FU),Doxorubicin (Adriamycin®), Prednisone, Tamoxifen (Nolvadex®), Paclitaxel(Taxol®), Albumin-bound paclitaxel (nab-paclitaxel, Abraxane®),Leucovorin, Thiotepa (Thioplex®), Anastrozole (Arimidex®), Docetaxel(Taxotere®), Vinorelbine (Navelbine®), Gemcitabine (Gemzar®), Ifosfamide(Ifex®), Pemetrexed (Alimta®), Topotecan, Melphalan (L-Pam®), Cisplatin(Cisplatinum®, Platinol®), Carboplatin (Paraplatin®), Oxaliplatin(Eloxatin®), Nedaplatin (Aqupla®), Triplatin, Lipoplatin (Nanoplatin®),Satraplatin, Picoplatin, Carmustine (BCNU; BiCNU®), Methotrexate(Folex®, Mexate®), Edatrexate, Mitomycin C (Mutamycin®), Mitoxantrone(Novantrone®), Vincristine (Oncovin®), Vinblastine (Velban®),Vinorelbine (Navelbine®), Vindesine (Eldisine®), Fenretinide, Topotecan,Irinotecan (Camptosar®), 9-amino-camptothecin [9-AC], Biantrazole,Losoxantrone, Etoposide, and Teniposide.

In one embodiment, the preferred combination partner for the IL-1βbinding antibody or a functional fragment thereof (e.g., canakinumab orgevokizumab) is a mitotic inhibitor, preferably docetaxel. In oneembodiment, the preferred combination partner for canakinumab is amitotic inhibitor, preferably docetaxel. In one embodiment, thepreferred combination partner for gevokizumab is a mitotic inhibitor,preferably docetaxel. In one embodiment said combination is used for thetreatment of CML.

In one embodiment, the preferred combination partner for the IL-1βbinding antibody or a functional fragment thereof (e.g., canakinumab orgevokizumab) is a platinum agent, preferably cisplatin. In oneembodiment, the preferred combination partner for canakinumab is aplatinum agent, preferably cisplatin. In one embodiment, the preferredcombination partner for gevokizumab is a platinum agent, preferablycisplatin. In one embodiment, the one or more chemotherapeutic agent isa platinum-based doublet chemotherapy (PT-DC).

Chemotherapy may comprise the administration of a single anti-canceragent (drug) or the administration of a combination of anti-canceragents (drugs), for example, one of the following, commonly administeredcombinations of: carboplatin and taxol; gemcitabine and cisplatin;gemcitabine and vinorelbine; gemcitabine and paclitaxel; cisplatin andvinorelbine; cisplatin and gemcitabine; cisplatin and paclitaxel(Taxol); cisplatin and docetaxel (Taxotere); cisplatin and etoposide;cisplatin and pemetrexed; carboplatin and vinorelbine; carboplatin andgemcitabine; carboplatin and paclitaxel (Taxol); carboplatin anddocetaxel (Taxotere); carboplatin and etoposide; carboplatin andpemetrexed. In one embodiment, the one or more chemotherapeutic agent isa platinum-based doublet chemotherapy (PT-DC).

Another class of chemotherapeutic agents are the inhibitors, especiallytyrosine kinase inhibitors, that specifically target growth promotingreceptors, especially VEGF-R, EGFR, PFGF-R and ALK, or their downstreammembers of the signalling transduction pathway, the mutation oroverproduction of which results in or contributes to the oncogenesis ofthe tumor at the site (targeted therapies). Exemplary of targetedtherapies drugs approved by the Food and Drug administration (FDA) forthe targeted treatment of lung cancer include but not limitedbevacizumab (Avastin®), crizotinib (Xalkori®), erlotinib (Tarceva®),gefitinib (Iressa®), afatinib dimaleate (Gilotrif®), ceritinib(LDK378/Zykadia™), everolimus (Afinitor®), ramucirumab (Cyramza®),osimertinib (Tagrisso™), necitumumab (Portrazza™), alectinib(Alecensa®), atezolizumab (Tecentriq™), brigatinib (Alunbrig™),trametinib (Mekinist®), dabrafenib (Tafinlar®), sunitinib (Sutent®) andcetuximab (Erbitux®).

In one embodiment the one or more chemotherapeutic agent to be combinedwith the IL-1β binding antibody or fragment thereof, suitablycanakinumab or gevokizumab, is the agent that is the standard of careagent for lung cancer, including NSCLC and SCLC. Standard of care, canbe found, for example from American Society of Clinical Oncology (ASCO)guideline on the systemic treatment of patients with stage IVnon-small-cell lung cancer (NSCLC) or American Society of ClinicalOncology (ASCO) guideline on Adjuvant Chemotherapy and AdjuvantRadiation Therapy for Stages I-IIIA Resectable Non-Small Cell LungCancer.

In one embodiment the one or more chemotherapeutic agent to be combinedwith the IL-1β binding antibody or fragment thereof, suitablycanakinumab or gevokizumab, is a platinum containing agent or aplatinum-based doublet chemotherapy (PT-DC). In one embodiment saidcombination is used for the treatment of lung cancer, especially NSCLC.In one embodiment one or more chemotherapeutic agent is a tyrosinekinase inhibitor. In one preferred embodiment said tyrosine kinaseinhibitor is a VEGF pathway inhibitor or an EGF pathway inhibitor. Inone embodiment the one or more chemotherapeutic agent is check-pointinhibitor, preferably pembrolizumab. In one embodiment said combinationis used for the treatment of lung cancer, especially NSCLC.

In one embodiment the one or more therapeutic agent to be combined withthe IL-1β binding antibody or fragment thereof, suitably canakinumab orgevokizumab, is a check-point inhibitor. In one further embodiment, saidcheck-point inhibitor is nivolumab. In one embodiment said check-pointinhibitor is pembrolizumab. In one further embodiment, said check-pointinhibitor is atezolizumab. In one further embodiment, said check-pointinhibitor is PDR-001 (spartalizumab). In one embodiment, saidcheck-point inhibitor is durvalumab. In one embodiment, said check-pointinhibitor is avelumab. Immunotherapies that target immune checkpoints,also known as checkpoint inhibitors, are currently emerging as keyagents in cancer therapy. The immune checkpoint inhibitor can be aninhibitor of the receptor or an inhibitor of the ligand. Examples of theinhibiting targets include but not limited to a co-inhibitory molecule(e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), a PD-L1inhibitor (e.g., an anti-PD-L1 antibody molecule), a PD-L2 inhibitor(e.g., an anti-PD-L2 antibody molecule), a LAG-3 inhibitor (e.g., ananti-LAG-3 antibody molecule), a TIM-3 inhibitor (e.g., an anti-TIM-3antibody molecule)), an activator of a co-stimulatory molecule (e.g., aGITR agonist (e.g., an anti-GITR antibody molecule)), a cytokine (e.g.,IL-15 complexed with a soluble form of IL-15 receptor alpha (IL-15Ra)),an inhibitor of cytotoxic T-lymphocyte-associated protein 4 (e.g., ananti-CTLA-4 antibody molecule) or any combination thereof.

PD-1 Inhibitors

In one aspect of the invention, the IL-1β inhibitor or a functionalfragment thereof is administered together with a PD-1 inhibitor. In onesome embodiment the PD-1 inhibitor is chosen from PDR001 (spartalizumab)(Novartis), Nivolumab (Bristol-Myers Squibb), Pembrolizumab (Merck &Co), Pidilizumab (CureTech), MEDI0680 (Medimmune), REGN2810 (Regeneron),TSR-042 (Tesaro), PF-06801591 (Pfizer), BGB-A317 (Beigene), BGB-108(Beigene), INCSHR1210 (Incyte), or AMP-224 (Amplimmune).

In one embodiment, the PD-1 inhibitor is an anti-PD-1 antibody. In oneembodiment, the PD-1 inhibitor is an anti-PD-1 antibody molecule asdescribed in US 2015/0210769, published on Jul. 30, 2015, entitled“Antibody Molecules to PD-1 and Uses Thereof,” incorporated by referencein its entirety.

In one embodiment, the anti-PD-1 antibody molecule comprises a VHcomprising the amino acid sequence of SEQ ID NO: 506 and a VL comprisingthe amino acid sequence of SEQ ID NO: 520. In one embodiment, theanti-PD-1 antibody molecule comprises a VH comprising the amino acidsequence of SEQ ID NO: 506 and a VL comprising the amino acid sequenceof SEQ ID NO: 516.

TABLE A  Amino acid and nucleotide sequences ofexemplary anti-PD-1 antibody molecules BAP049-Clone-B HC SEQ ID NO: 506VH EVQLVQSGAEVKKPGESLRISCKGSGYT FTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELS SLRSEDTAVYYCTRWTTGTGAYWGQGTT VTVSSBAP049-Clone-B LC SEQ ID NO: 516 VL EIVLTQSPATLSLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGKAPKLLIYW ASTRESGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQNDYSYPYTFGQGTKVEI K BAP049-Clone-E HC SEQ ID NO: 506 VHEVQLVQSGAEVKKPGESLRISCKGSGYT FTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELS SLRSEDTAVYYCTRWTTGTGAYWGQGTT VTVSSBAP049-Clone-E LC SEQ ID NO: 520 VL EIVLTQSPATLSLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYW ASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEI K

In one embodiment, the anti-PD-1 antibody is spartalizumab.

In one embodiment, the anti-PD-1 antibody is Nivolumab.

In one embodiment, the anti-PD-1 antibody molecule is Pembrolizumab.

In one embodiment, the anti-PD-1 antibody molecule is Pidilizumab.

In one embodiment, the anti-PD-1 antibody molecule is MEDI0680(Medimmune), also known as AMP-514. MEDI0680 and other anti-PD-1antibodies are disclosed in U.S. Pat. No. 9,205,148 and WO 2012/145493,incorporated by reference in their entirety. Other exemplary anti-PD-1molecules include REGN2810 (Regeneron), PF-06801591 (Pfizer),BGB-A317/BGB-108 (Beigene), INCSHR1210 (Incyte) and TSR-042 (Tesaro).

Further known anti-PD-1 antibodies include those described, e.g., in WO2015/112800, WO 2016/092419, WO 2015/085847, WO 2014/179664, WO2014/194302, WO 2014/209804, WO 2015/200119, U.S. Pat. Nos. 8,735,553,7,488,802, 8,927,697, 8,993,731, and 9,102,727, incorporated byreference in their entirety.

In one embodiment, the anti-PD-1 antibody is an antibody that competesfor binding with, and/or binds to the same epitope on PD-1 as, one ofthe anti-PD-1 antibodies described herein.

In one embodiment, the PD-1 inhibitor is a peptide that inhibits thePD-1 signaling pathway, e.g., as described in U.S. Pat. No. 8,907,053,incorporated by reference in its entirety. In one embodiment, the PD-1inhibitor is an immunoadhesin (e.g., an immunoadhesin comprising anextracellular or PD-1 binding portion of PD-L1 or PD-L2 fused to aconstant region (e.g., an Fc region of an immunoglobulin sequence). Inone embodiment, the PD-1 inhibitor is AMP-224 (B7-DCIg (Amplimmune),e.g., disclosed in WO 2010/027827 and WO 2011/066342, incorporated byreference in their entirety).

PD-L1 Inhibitors

In one aspect of the invention, the IL-1β inhibitor or a functionalfragment thereof is administered together with a PD-L1 inhibitor. Insome embodiments, the PD-L1 inhibitor is chosen from FAZ053 (Novartis),Atezolizumab (Genentech/Roche), Avelumab (Merck Serono and Pfizer),Durvalumab (MedImmune/AstraZeneca), or BMS-936559 (Bristol-MyersSquibb).

In one embodiment, the PD-L1 inhibitor is an anti-PD-L1 antibodymolecule. In one embodiment, the PD-L1 inhibitor is an anti-PD-L1antibody molecule as disclosed in US 2016/0108123, published on Apr. 21,2016, entitled “Antibody Molecules to PD-L1 and Uses Thereof,”incorporated by reference in its entirety.

In one embodiment, the anti-PD-L1 antibody molecule comprises a VHcomprising the amino acid sequence of SEQ ID NO: 606 and a VL comprisingthe amino acid sequence of SEQ ID NO: 616. In one embodiment, theanti-PD-L1 antibody molecule comprises a VH comprising the amino acidsequence of SEQ ID NO: 620 and a VL comprising the amino acid sequenceof SEQ ID NO: 624.

TABLE B  Amino acid and nucleotide sequences of exemplaryanti-PD-L1 antibody molecules BAP058-Clone O HC SEQ ID NO: 606 VHEVQLVQSGAEVKKPGATVKISCKVSGYTF TSYWMYWVRQARGQRLEWIGRIDPNSGSTKYNEKFKNRFTISRDNSKNTLYLQMNSLR AEDTAVYYCARDYRKGLYAMDYWGQGTTV TVSSBAP058-Clone O LC SEQ ID NO: 616 VL AIQLTQSPSSLSASVGDRVTITCKASQDVGTAVAWYLQKPGQSPQLLIYWASTRHTGV PSRFSGSGSGTDFTFTISSLEAEDAATYYCQQYNSYPLTFGQGTKVEIK BAP058-Clone N HC SEQ ID NO: 620 VHEVQLVQSGAEVKKPGATVKISCKVSGYTF TSYWMYWVRQATGQGLEWMGRIDPNSGSTKYNEKFKNRVTITADKSTSTAYMELSSLR SEDTAVYYCARDYRKGLYAMDYWGQGTTV TVSSBAP058-Clone N LC SEQ ID NO: 624 VL DVVMTQSPLSLPVTLGQPASISCKASQDVGTAVAWYQQKPGQAPRLLIYWASTRHTGV PSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYNSYPLTFGQGTKVEIK

In one embodiment, the anti-PD-L1 antibody molecule is Atezolizumab(Genentech/Roche), also known as MPDL3280A, RG7446, RO5541267,YW243.55.S70, or TECENTRIQ™. Atezolizumab and other anti-PD-L1antibodies are disclosed in U.S. Pat. No. 8,217,149, incorporated byreference in its entirety.

In one embodiment, the anti-PD-L1 antibody molecule is Avelumab (MerckSerono and Pfizer), also known as MSB0010718C. Avelumab and otheranti-PD-L1 antibodies are disclosed in WO 2013/079174, incorporated byreference in its entirety.

In one embodiment, the anti-PD-L1 antibody molecule is Durvalumab(MedImmune/AstraZeneca), also known as MEDI4736. Durvalumab and otheranti-PD-L1 antibodies are disclosed in U.S. Pat. No. 8,779,108,incorporated by reference in its entirety.

In one embodiment, the anti-PD-L1 antibody molecule is BMS-936559(Bristol-Myers Squibb), also known as MDX-1105 or 12A4. BMS-936559 andother anti-PD-L1 antibodies are disclosed in U.S. Pat. No. 7,943,743 andWO 2015/081158, incorporated by reference in their entirety.

Further known anti-PD-L1 antibodies include those described, e.g., in WO2015/181342, WO 2014/100079, WO 2016/000619, WO 2014/022758, WO2014/055897, WO 2015/061668, WO 2013/079174, WO 2012/145493, WO2015/112805, WO 2015/109124, WO 2015/195163, U.S. Pat. Nos. 8,168,179,8,552,154, 8,460,927, and 9,175,082, incorporated by reference in theirentirety.

In one embodiment, the anti-PD-L1 antibody is an antibody that competesfor binding with, and/or binds to the same epitope on PD-L1 as, one ofthe anti-PD-L1 antibodies described herein.

LAG-3 Inhibitors

In one aspect of the invention, the IL-1β inhibitor or a functionalfragment thereof is administered together with a LAG-3 inhibitor. Insome embodiments, the LAG-3 inhibitor is chosen from LAG525 (Novartis),BMS-986016 (Bristol-Myers Squibb), TSR-033 (Tesaro), IMP731 orGSK2831781 and IMP761 (Prima BioMed).

In one embodiment, the LAG-3 inhibitor is an anti-LAG-3 antibodymolecule. In one embodiment, the LAG-3 inhibitor is an anti-LAG-3antibody molecule as disclosed in US 2015/0259420, published on Sep. 17,2015, entitled “Antibody Molecules to LAG-3 and Uses Thereof,”incorporated by reference in its entirety.

In one embodiment, the anti-LAG-3 antibody molecule comprises a VHcomprising the amino acid sequence of SEQ ID NO: 706 and a VL comprisingthe amino acid sequence of SEQ ID NO: 718. In one embodiment, theanti-LAG-3 antibody molecule comprises a VH comprising the amino acidsequence of SEQ ID NO: 724 and a VL comprising the amino acid sequenceof SEQ ID NO: 730.

TABLE C  Amino acid and nucleotide sequences of exemplaryanti-LAG-3 antibody molecules BAP050-Clone I HC SEQ ID NO: 706 VHQVQLVQSGAEVKKPGASVKVSCKASGFTLT NYGMNWVRQARGQRLEWIGWINTDTGEPTYADDFKGRFVFSLDTSVSTAYLQISSLKAED TAVYYCARNPPYYYGTNNAEAMDYWGQGTT VTVSSBAP050-Clone I LC SEQ ID NO: 718 VL DIQMTQSPSSLSASVGDRVTITCSSSQDISNYLNWYLQKPGQSPQLLIYYTSTLHLGVPS RFSGSGSGTEFTLTISSLQPDDFATYYCQQYYNLPWTFGQGTKVEIK BAP050-Clone J HC SEQ ID NO: 724 VHQVQLVQSGAEVKKPGASVKVSCKASGFTLT NYGMNWVRQAPGQGLEWMGWINTDTGEPTYADDFKGRFVFSLDTSVSTAYLQISSLKAED TAVYYCARNPPYYYGTNNAEAMDYWGQGTT VTVSSBAP050-Clone J LC SEQ ID NO: 730 VL DIQMTQSPSSLSASVGDRVTITCSSSQDISNYLNWYQQKPGKAPKLLIYYTSTLHLGIPP RFSGSGYGTDFTLTINNIESEDAAYYFCQQYYNLPWTFGQGTKVEIK

In one embodiment, the anti-LAG-3 antibody molecule is BMS-986016(Bristol-Myers Squibb), also known as BMS986016. BMS-986016 and otheranti-LAG-3 antibodies are disclosed in WO 2015/116539 and U.S. Pat. No.9,505,839, incorporated by reference in their entirety. In oneembodiment, the anti-LAG-3 antibody molecule comprises one or more ofthe CDR sequences (or collectively all of the CDR sequences), the heavychain or light chain variable region sequence, or the heavy chain orlight chain sequence of BMS-986016, e.g., as disclosed in Table D.

In one embodiment, the anti-LAG-3 antibody molecule is IMP731 orGSK2831781 (GSK and Prima BioMed). IMP731 and other anti-LAG-3antibodies are disclosed in WO 2008/132601 and U.S. Pat. No. 9,244,059,incorporated by reference in their entirety. In one embodiment, theanti-LAG-3 antibody molecule comprises one or more of the CDR sequences(or collectively all of the CDR sequences), the heavy chain or lightchain variable region sequence, or the heavy chain or light chainsequence of IMP731, e.g., as disclosed in Table D.

Further known anti-LAG-3 antibodies include those described, e.g., in WO2008/132601, WO 2010/019570, WO 2014/140180, WO 2015/116539, WO2015/200119, WO 2016/028672, U.S. Pat. Nos. 9,244,059, 9,505,839,incorporated by reference in their entirety.

In one embodiment, the anti-LAG-3 antibody is an antibody that competesfor binding with, and/or binds to the same epitope on LAG-3 as, one ofthe anti-LAG-3 antibodies described herein.

In one embodiment, the anti-LAG-3 inhibitor is a soluble LAG-3 protein,e.g., IMP321 (Prima BioMed), e.g., as disclosed in WO 2009/044273,incorporated by reference in its entirety.

TABLE D  Amino acid sequences of exemplary anti-LAG-3 antibody moleculesBMS-986016 SEQ ID NO: 762 Heavy QVQLQQWGAGLLKPSETLSLTCAVYGGSFSDYYWNWIRQPPGKG chainLEWIGEINHRGSTNSNPSLKSRVTLSLDTSKNQFSLKLRSVTAADTAVYYCAFGYSDYEYNWFDPWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 763 Light EIVLTQSPATLSLSPGERATLSCRASQSISSYLAWYQQKPGQAPRLL chainIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPLTFGQGTNLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC IMP731 SEQ ID NO: 764 HeavyQVQLKESGPGLVAPSQSLSITCTVSGFSLTAYGVNWVRQPPGKGLE chainWLGMIWDDGSTDYNSALKSRLSISKDNSKSQVFLKMNSLQTDDTARYYCAREGDVAFDYWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL SEQ ID NO: 765 Light TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK chainDIVMTQSPSSLAVSVGQKVTMSCKSSQSLLNGSNQKNYLAWYQQKPGQSPKLLVYFASTRDSGVPDRFIGSGSGTDFTLTISSVQAEDLADYFCLQHFGTPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

TIM-3 Inhibitors

In one aspect of the invention, the IL-1β inhibitor or a functionalfragment thereof is administered together with a TIM-3 inhibitor. Insome embodiments, the TIM-3 inhibitor is MGB453 (Novartis) or TSR-022(Tesaro).

In one embodiment, the TIM-3 inhibitor is an anti-TIM-3 antibodymolecule. In one embodiment, the TIM-3 inhibitor is an anti-TIM-3antibody molecule as disclosed in US 2015/0218274, published on Aug. 6,2015, entitled “Antibody Molecules to TIM-3 and Uses Thereof,”incorporated by reference in its entirety.

In one embodiment, the anti-TIM-3 antibody molecule comprises a VHcomprising the amino acid sequence of SEQ ID NO: 806 and a VL comprisingthe amino acid sequence of SEQ ID NO: 816. In one embodiment, theanti-TIM-3 antibody molecule comprises a VH comprising the amino acidsequence of SEQ ID NO: 822 and a VL comprising the amino acid sequenceof SEQ ID NO: 826.

The antibody molecules described herein can be made by vectors, hostcells, and methods described in US 2015/0218274, incorporated byreference in its entirety.

TABLE E  Amino acid and nucleotide sequences of exemplaryanti-TIM-3 antibody molecules ABTIM3-hum11 SEQ ID NO: 806 VHQVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIYPGNGDTSYNQKFKGRVTITADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSS SEQ ID NO: 816 VLAIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPKLLIYAASNVESGVPSRFSGSGSGTDFTLTISSLQPE DFATYFCQQSRKDPSTFGGGTKVEIKABTIM3-hum03 SEQ ID NO: 822 VH QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNM HWVRQAPGQGLEWIGDIYPGQGDTSYNQKFKGRATMTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSS SEQ ID NO: 826 VLDIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLIYAASNVESGVPDRFSGSGSGTDFTLTISSLQA EDVAVYYCQQSRKDPSTFGGGTKVEIK

In one embodiment, the anti-TIM-3 antibody molecule is TSR-022(AnaptysBio/Tesaro). In one embodiment, the anti-TIM-3 antibody moleculecomprises one or more of the CDR sequences (or collectively all of theCDR sequences), the heavy chain or light chain variable region sequence,or the heavy chain or light chain sequence of TSR-022. In oneembodiment, the anti-TIM-3 antibody molecule comprises one or more ofthe CDR sequences (or collectively all of the CDR sequences), the heavychain or light chain variable region sequence, or the heavy chain orlight chain sequence of APE5137 or APE5121, e.g., as disclosed in TableF. APE5137, APE5121, and other anti-TIM-3 antibodies are disclosed in WO2016/161270, incorporated by reference in its entirety.

In one embodiment, the anti-TIM-3 antibody molecule is the antibodyclone F38-2E2. In one embodiment, the anti-TIM-3 antibody moleculecomprises one or more of the CDR sequences (or collectively all of theCDR sequences), the heavy chain or light chain variable region sequence,or the heavy chain or light chain sequence of F38-2E2.

Further known anti-TIM-3 antibodies include those described, e.g., in WO2016/111947, WO 2016/071448, WO 2016/144803, U.S. Pat. Nos. 8,552,156,8,841,418, and 9,163,087, incorporated by reference in their entirety.

In one embodiment, the anti-TIM-3 antibody is an antibody that competesfor binding with, and/or binds to the same epitope on TIM-3 as, one ofthe anti-TIM-3 antibodies described herein.

TABLE F  Amino acid sequences of exemplary anti-TIM-3 antibody moleculesAPE5137 SEQ ID NO: 830 VHEVQLLESGGGLVQPGGSLRLSCAAASGFTFSSYDMSWVRQAPGKGLDWVSTISGGGTYTYYQDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC ASMDYWGQGTTVTVSSASEQ ID NO: 831 VL DIQMTQSPSSLSASVGDRVTITCRASQSIRRYLNWYHQKPGKAPKLLIYGASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFAVYYCQQSHSAPLTFGG GTKVEIKR APE5121SEQ ID NO: 832 VH EVQVLESGGGLVQPGGSLRLYCVASGFTFSGSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKKYYVGPADYWGQGTLVTVSSG SEQ ID NO: 833 VLDIVMTQSPDSLAVSLGERATINCKSSQSVLYSSNNKNYLAWYQHKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYS SPLTFGGGTKIEVK

GITR Agonists

In one aspect of the invention, the IL-1β inhibitor or a functionalfragment thereof is administered together with a GITR agonist. In someembodiments, the GITR agonist is GWN323 (NVS), BMS-986156, MK-4166 orMK-1248 (Merck), TRX518 (Leap Therapeutics), INCAGN1876 (Incyte/Agenus),AMG 228 (Amgen) or INBRX-110 (Inhibrx).

In one embodiment, the GITR agonist is an anti-GITR antibody molecule.In one embodiment, the GITR agonist is an anti-GITR antibody molecule asdescribed in WO 2016/057846, published on Apr. 14, 2016, entitled“Compositions and Methods of Use for Augmented Immune Response andCancer Therapy,” incorporated by reference in its entirety.

In one embodiment, the anti-GITR antibody molecule comprises a VHcomprising the amino acid sequence of SEQ ID NO: 901 and a VL comprisingthe amino acid sequence of SEQ ID NO: 902.

TABLE G  Amino acid and nucleotide sequences of exemplaryanti-GITR antibody molecule MAB7 SEQ ID VHEVQLVESGGGLVQSGGSLRLSCAASGFSLSSYGVDWV NO: 901RQAPGKGLEWVGVIWGGGGTYYASSLMGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARHAYGHDGGFAMDYW GQGTLVTVSS SEQ ID  VLEIVMTQSPATLSVSPGERATLSCRASESVSSNVAWYQ NO: 902QRPGQAPRLLIYGASNRATGIPARFSGSGSGTDFTLT ISRLEPEDFAVYYCGQSYSYPFTFGQGTKLEIK

In one embodiment, the anti-GITR antibody molecule is BMS-986156(Bristol-Myers Squibb), also known as BMS 986156 or BMS986156.BMS-986156 and other anti-GITR antibodies are disclosed, e.g., in U.S.Pat. No. 9,228,016 and WO 2016/196792, incorporated by reference intheir entirety. In one embodiment, the anti-GITR antibody moleculecomprises one or more of the CDR sequences (or collectively all of theCDR sequences), the heavy chain or light chain variable region sequence,or the heavy chain or light chain sequence of BMS-986156, e.g., asdisclosed in Table H.

In one embodiment, the anti-GITR antibody molecule is MK-4166 or MK-1248(Merck). MK-4166, MK-1248, and other anti-GITR antibodies are disclosed,e.g., in U.S. Pat. No. 8,709,424, WO 2011/028683, WO 2015/026684, andMahne et al. Cancer Res. 2017; 77(5):1108-1118, incorporated byreference in their entirety.

In one embodiment, the anti-GITR antibody molecule is TRX518 (LeapTherapeutics). TRX518 and other anti-GITR antibodies are disclosed,e.g., in U.S. Pat. Nos. 7,812,135, 8,388,967, 9,028,823, WO 2006/105021,and Ponte J et al. (2010) Clinical Immunology; 135:S96, incorporated byreference in their entirety.

In one embodiment, the anti-GITR antibody molecule is INCAGN1876(Incyte/Agenus). INCAGN1876 and other anti-GITR antibodies aredisclosed, e.g., in US 2015/0368349 and WO 2015/184099, incorporated byreference in their entirety.

In one embodiment, the anti-GITR antibody molecule is AMG 228 (Amgen).AMG 228 and other anti-GITR antibodies are disclosed, e.g., in U.S. Pat.No. 9,464,139 and WO 2015/031667, incorporated by reference in theirentirety.

In one embodiment, the anti-GITR antibody molecule is INBRX-110(Inhibrx). INBRX-110 and other anti-GITR antibodies are disclosed, e.g.,in US 2017/0022284 and WO 2017/015623, incorporated by reference intheir entirety.

In one embodiment, the GITR agonist (e.g., a fusion protein) is MEDI1873 (MedImmune), also known as MEDI1873. MEDI 1873 and other GITRagonists are disclosed, e.g., in US 2017/0073386, WO 2017/025610, andRoss et al. Cancer Res 2016; 76(14 Suppl): Abstract nr 561, incorporatedby reference in their entirety. In one embodiment, the GITR agonistcomprises one or more of an IgG Fc domain, a functional multimerizationdomain, and a receptor binding domain of a glucocorticoid-induced TNFreceptor ligand (GITRL) of MEDI 1873.

Further known GITR agonists (e.g., anti-GITR antibodies) include thosedescribed, e.g., in WO 2016/054638, incorporated by reference in itsentirety.

In one embodiment, the anti-GITR antibody is an antibody that competesfor binding with, and/or binds to the same epitope on GITR as, one ofthe anti-GITR antibodies described herein.

In one embodiment, the GITR agonist is a peptide that activates the GITRsignaling pathway. In one embodiment, the GITR agonist is animmunoadhesin binding fragment (e.g., an immunoadhesin binding fragmentcomprising an extracellular or GITR binding portion of GITRL) fused to aconstant region (e.g., an Fc region of an immunoglobulin sequence).

TABLE H  Amino acid sequence of exemplary anti-GITR antibody moleculesBMS-986156 SEQ ID  VH QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGM NO: 920HWVRQAPGKGLEWVAVIWYEGSNKYYADSVKGRF TISRDNSKNTLYLQMNSLRAEDTAVYYCARGGSMVRGDYYYGMDVWGQGTTVTVSS SEQ ID  VL AIQLTQSPSSLSASVGDRVTITCRASQGISSALANO: 921 WYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPYTFGQGT KLEIK

IL15/IL-15Ra Complexes

In one aspect of the invention, the IL-1β inhibitor or a functionalfragment thereof is administered together with an IL-15/IL-15Ra complex.In some embodiments, the IL-15/IL-15Ra complex is chosen from NIZ985(Novartis), ATL-803 (Altor) or CYP0150 (Cytune).

In one embodiment, the IL-15/IL-15Ra complex comprises human IL-15complexed with a soluble form of human IL-15Ra. The complex may compriseIL-15 covalently or noncovalently bound to a soluble form of IL-15Ra. Ina particular embodiment, the human IL-15 is noncovalently bonded to asoluble form of IL-15Ra. In a particular embodiment, the human IL-15 ofthe composition comprises an amino acid sequence of SEQ ID NO: 1001 inTable I and the soluble form of human IL-15Ra comprises an amino acidsequence of SEQ ID NO:1002 in Table I, as described in WO 2014/066527,incorporated by reference in its entirety. The molecules describedherein can be made by vectors, host cells, and methods described in WO2007/084342, incorporated by reference in its entirety.

TABLE I  Amino acid and nucleotide sequences ofexemplary IL-153L-15Ra complexes NIZ985 SEQ ID Human NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPS NO: 1001 IL-15CKVTAMKCFLLELQVISLESGDASIHDTVENLII LANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS SEQ ID Human  ITCPPPMSVEHADIWVKSYSLYSRERYICNSGFK NO: 1002Soluble RKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPA IL-15RaLVHQRPAPPSTVTTAGVTPQPESLSPSGKEPAAS SPSSNNTAATTAAIVPGSQLMPSKSPSTGTTEISSHESSHGTPSQTTAKNWELTASASHQPPGVYPQG

In one embodiment, the IL-15/IL-15Ra complex is ALT-803, anIL-15/IL-15Ra Fc fusion protein (IL-15N72D:IL-15RaSu/Fc solublecomplex). ALT-803 is disclosed in WO 2008/143794, incorporated byreference in its entirety. In one embodiment, the IL-15/IL-15Ra Fcfusion protein comprises the sequences as disclosed in Table J.

In one embodiment, the IL-15/IL-15Ra complex comprises IL-15 fused tothe sushi domain of IL-15Ra (CYP0150, Cytune). The sushi domain ofIL-15Ra refers to a domain beginning at the first cysteine residue afterthe signal peptide of IL-15Ra, and ending at the fourth cysteine residueafter said signal peptide. The complex of IL-15 fused to the sushidomain of IL-15Ra is disclosed in WO 2007/04606 and WO 2012/175222,incorporated by reference in their entirety. In one embodiment, theIL-15/IL-15Ra sushi domain fusion comprises the sequences as disclosedin Table J.

TABLE J  Amino acid sequences of other exemplary IL-15/IL-15Ra complexesALT-803 (Altor) SEQ ID NO: IL-15N72DNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTA 1003MKCFLLELQVISLESGDASIHDTVENLIILANDSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS SEQ ID NO: IL-15RaSu/FcITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVL 1004NKATNVAHWTTPSLKCIREPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPGKIL-15/IL-15Ra sushi domain fusion (Cytune) SEQ ID Human IL-15NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLE NO: 1005LQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEXK NIKEFLQSFVHIVQMFINTSWhere X is E or K SEQ ID Human IL-ITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVL NO: 1006 15Ra sushi NKATNVAHWTTPSLKCIRDPALVHQRPAPP and hinge domains

CTLA-4 Inhibitors

In one aspect of the invention, the IL-1β inhibitor or a functionalfragment thereof is administered together with an inhibitor of CTLA-4.In some embodiments, the CTLA-4 inhibitor is an anti-CTLA-4 antibody orfragment thereof. Exemplary anti-CTLA-4 antibodies include Tremelimumab(formerly ticilimumab, CP-675,206); and Ipilimumab (MDX-010, Yervoy®).In one embodiment, the present invention provides an IL-1β antibody or afunctional fragment thereof (e.g., canakinumab or gevokizumab) for usein the treatment cancers having at least partial inflammatory bases,e.g., CML, wherein said IL-1β antibody or a functional fragment thereofis administered in combination with one or more chemotherapeutic agent,wherein said one or more chemotherapeutic agent is a check pointinhibitor, preferably selected from the group consisting of nivolumab,pembrolizumab, atezolizumab, avelumab, durvalumab, PDR-001(spartalizumab) and Ipilimumab. In one embodiment the one or morechemotherapeutic agent is a PD-1 or PD-L-1 inhibitor, preferablyselected from the group consisting of nivolumab, pembrolizumab,atezolizumab, avelumab, durvalumab, PDR-001 (spartalizumab), furtherpreferably pembrolizumab. In one further embodiment, the IL-1β antibodyor a functional fragment thereof is administered at the same time of thePD-1 or PD-L1 inhibitor.

In one embodiment said patient has a tumor that has high PD-L1expression [Tumor Proportion Score (TPS) ≥50%)] as determined by anFDA-approved test, with or without EGFR or ALK genomic tumoraberrations. In one embodiment said patient has tumor that has PD-L1expression (TPS ≥1%) as determined by an FDA-approved test.

In one embodiment the one or more therapeutic agents is lacnotuzumab. Inone embodiment the one or more therapeutic agents further include acheck point inhibitor, suitably a check point inhibitor, suitablyselected from pembrolizumab, nivolumab, spartalizumab, atezolizumab,avelumab, ipilimumab, durvalumab. In one embodiment the cancer is CML.Lacnotuzumab is administered at a dose of about 3 mg/kg, about 5 mg/kg,about 7.5 mg/kg or about 10 mg/kg body weight, preferably about every 3weeks or about every 4 weeks.

In one embodiment, the one or more chemotherapeutic agents ismidostaurin (Rydapt®). In one embodiment the one or morechemotherapeutic agents further include cytarabine and daunorubicin,preferably in combination with standard cytarabine and daunorubicininduction and cytarabine consolidation. In one embodiment, midostaurinis administered about 50 mg orally twice daily with food. In a preferredembodiment, midostaurin is administered about 50 mg orally twice dailywith food on Days 8 to 21 of each cycle of induction with cytarabine anddaunorubicin and on Days 8 to 21 of each cycle of consolidation withhigh-dose cytarabine. In one embodiment, canakinumab is administeredabout 200 mg about every 4 weeks, in combination with ribociclib. In oneembodiment, gevokizumab is administered about 30-120 mg about every 4weeks, in combination with ribociclib.

In one embodiment, the one or more chemotherapeutic agents is5-bromo-2,6-di-(1H-pyrazol-1-yl)pyrimidine-4-amine or a pharmaceuticallyacceptable salt thereof (the compound described in Example 1 in the PCTpublication WO 2011/121418, which is hereby incorporated by reference inits entirety. In one embodiment, the cancer is CML.

In one embodiment, the one or more chemotherapeutic agents is4-[2-((1R,2R)-2-Hydroxy-cyclohexylamino)-benzothiazol-6-yloxy]-pyridine-2-carboxylicacid methylamide or a pharmaceutically acceptable salt thereof (compound157 in the PCT publication WO 2007/121484 A2, which is herebyincorporated by reference in its entirety). In one embodiment, thecancer is CML.

In one embodiment, the one or more therapeutic agents is a TGF-betainhibitor, preferably NIS793.

The heavy chain variable region of NIS793 has the amino acid sequenceof:

(SEQ ID NO: 6 in WO 2012/167143)QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGLWEVRALPSVYWGQGTLVTVSS.The light chain variable region of NIS793 has the amino acid sequenceof:

(SEQ ID NO: 8 in WO 2012/167143)SYELTQPPSVSVAPGQTARITCGANDIGSKSVHWYQQKAGQAPVLVVSEDIIRPSGIPERISGSNSGNTATLTISRVEAGDEADYYCQVWDRDSDQYVFG TGTKVTVLG.

NIS793 is a fully human monoclonal antibody that specifically binds andneutralizes TGF-beta 1 and 2 ligands. In one embodiment, the one or moretherapeutic agents further includes one PD-1 or PD-L1 inhibitor,suitably selected from selected from pembrolizumab, nivolumab,spartalizumab, atezolizumab, avelumab, ipilimumab, durvalumab. suitablypembrolizumab, suitably spartalizumab. In one embodiment, the cancer isCML.

In one embodiment, the one or more chemotherapeutic agents is ribociclibor any pharmaceutical salt thereof. In one embodiment, the cancer isCML.

In one embodiment, ribociclib is administered at a dose of about 600 mgdaily for about 21 consecutive days followed by about 7 days offtreatment resulting in an about 28-day full cycle. In one embodiment,canakinumab is administered 200 mg every 4 weeks, in combination withribociclib. In one embodiment, gevokizumab is administered about 30-120mg about every 4 weeks, in combination with ribociclib.

The term “in combination with” is understood as the two or more drugsare administered subsequently or simultaneously. Alternatively, the term“in combination with” is understood that two or more drugs areadministered in the manner that the effective therapeutic concentrationof the drugs are expected to be overlapping for a majority of the periodof time within the patient's body. The DRUG of the invention and one ormore combination partner (e.g., another drug, also referred to as“therapeutic agent” or “co-agent”) may be administered independently atthe same time or separately within time intervals, especially wherethese time intervals allow that the combination partners show acooperative, e.g., synergistic effect. The terms “co-administration” or“combined administration” or the like as utilized herein are meant toencompass administration of the selected combination partner to a singlesubject in need thereof (e.g., a patient), and are intended to includetreatment regimens in which the agents are not necessarily administeredby the same route of administration or at the same time. The drugadministered to a patient as separate entities either simultaneously,concurrently or sequentially with no specific time limits, wherein suchadministration provides therapeutically effective levels of the twocompounds in the body of the patient and the treatment regimen willprovide beneficial effects of the drug combination in treating theconditions or disorders described herein. The latter also applies tococktail therapy, e.g., the administration of three or more activeingredients.

In one embodiment, the present invention provides an IL-1β antibody or afunctional fragment thereof, suitably canakinumab or a functionalfragment thereof or gevokizumab or a functional fragment thereof, foruse as the first line treatment of cancer having at least a partialinflammatory basis, e.g., CML. The term “first line treatment” meanssaid patient is given the IL-1β antibody or a functional fragmentthereof before the patient develops resistance to one or more otherchemotherapeutic agent. Preferably one or more other chemotherapeuticagent is a platinum-based mono or combination therapy, a targetedtherapy, such a tyrosine inhibitor therapy, a checkpoint inhibitortherapy or any combination thereof. As first line treatment, the IL-1βantibody or a functional fragment thereof, such as canakinumab orgevokizumab, can be administered to patient as monotherapy or preferablyin combination with an check point inhibitor, particularly a PD-1 orPD-L1 inhibitor, preferably pembrolizumab, with or without one or moresmall molecule chemotherapeutic agent. In one embodiment as first linetreatment, the IL-1β antibody or a functional fragment thereof, such ascanakinumab or gevokizumab, can be administered to patient incombination with the standard of care therapy for that cancer having atleast partial inflammatory basis, e.g., CML.

In one embodiment, the present invention provides an IL-1β antibody or afunctional fragment thereof, suitably canakinumab or a functionalfragment thereof or gevokizumab or a functional fragment thereof, foruse as the second or third line treatment of cancer having at least apartial inflammatory basis. The term “the second or third linetreatment” means IL-1β antibody or a functional fragment thereof isadministered to a patient with cancer progression on or after one ormore other therapeutic agent treatment, especially disease progressionon or after FDA-approved first line therapy for the cancer having atleast a partial inflammatory basis. Preferably one or more othertherapeutic agent is a platinum-based mono or combination therapy, atargeted therapy, such a tyrosine inhibitor therapy, a checkpointinhibitor therapy or any combination thereof. As the second or thirdline treatment, the IL-1β antibody or a functional fragment thereof canbe administered to the patient as monotherapy or preferably incombination with one or more therapeutic agent, including thecontinuation of the early treatment with the same one or moretherapeutic agent.

For use as the second or third line treatment, the IL-1β antibody or afunctional fragment thereof, such as canakinumab or gevokizumab, can beadministered to patient as monotherapy or preferably in combination witha check-point inhibitor, particularly a PD-1 or PD-L1 inhibitor,particularly atezolizumab, with or without one or more small moleculechemotherapeutic agent.

Exemplar of Cancers to be Treated According to the Present Invention

In one aspect the present invention provides an IL-1β binding antibodyor a functional fragment thereof, suitably gevokizumab or a functionalfragment thereof, suitably canakinumab or a functional fragment thereof,alone or in combination, for use in the treatment of cancer having atleast partial inflammatory basis, wherein said cancer is CML. In oneaspect the present invention provides an IL-1β binding antibody or afunctional fragment thereof for use in the treatment of CML. The term“Chronic myeloid leukemia (CML)” (also known as chronic myelogenousleukemia) as used herein refers to a cancer of the white blood cells. Itis a form of leukemia characterized by the increased and unregulatedgrowth of myeloid cells in the bone marrow and the accumulation of thesecells in the blood. CML is a clonal bone marrow stem cell disorder inwhich a proliferation of mature granulocytes (neutrophils, eosinophilsand basophils) and their precursors is found. It is a type ofmyeloproliferative neoplasm associated with a characteristic chromosomaltranslocation called the Philadelphia chromosome.

CML is a clonal disease that originates from a single transformedhematopoietic stem cell (HSC) or multipotent progenitor cell (MPP)harboring the Philadelphia translocation t(9/22). The expression of thegene product of this translocation, the fusion oncogene BCR-ABL, inducesmolecular changes which result in expansion of the malignanthematopoiesis including the leukemic stem cell (LSC) pool and theoutgrowth and suppression of non-malignant hematopoiesis (Stam et al.,Mol Cell Biol. 1987, 7:1955-60). Myeloid cells (granulocytes, monocytes,megakaryocytes, erythrocytes), but also B- and T-cells express BCR-ABL,indicating the MPP or HSC as the start point of the disease (Fialkow etal., J. Clin. Invest. 1978, 62:815-23; Takahashi et al., Blood 1998,92:4758-63). In contrast to oncogenes causing AML, like MOZ-TIF2 orMLL-ENL, BCR-ABL does not confer self-renewal properties to committedprogenitor cells, but rather utilizes and enhances the self-renewalproperties of existing self-renewing cells, like HSCs or MPPs. Duringthe course of the disease, the leukemic stem cell pool expands and inthe final stage, the blast crisis, nearly all CD34⁺CD38⁻ cells carry thePhiladelphia translocation.

CML is often divided into three phases based on clinical characteristicsand laboratory findings. In the absence of intervention, CML typicallybegins in the chronic phase, and over the course of several yearsprogresses to an accelerated phase and ultimately to a blast crisis.Blast crisis is the terminal phase of CML and clinically behaves like anacute leukemia. Drug treatment will usually stop this progression ifstarted early. One of the drivers of the progression from chronic phasethrough acceleration and blast crisis is the acquisition of newchromosomal abnormalities (in addition to the Philadelphia chromosome).Some patients may already be in the accelerated phase or blast crisis bythe time they are diagnosed.

The term CML as used herein includes all the phases, e.g., chronicphase, accelerated phase, and blast crisis.

In one embodiment, the present invention provides DRUG of the invention,preferably canakinumab or gevokizumab, for use in the treatment of CML.The DRUG of the invention may be used in combination with one or moretherapeutic agent, e.g., chemotherapeutic agent or a check pointinhibitor. In one embodiment the therapeutic agent is the standard ofcare agent for CML. For example, DRUG of the invention in combinationwith Hh antagonists may be administered adjunctively with any of thetreatment modalities, such as chemotherapy, radiation, and/or surgery.In one embodiment, the DRUG of the invention can be used in combinationwith one or more chemotherapeutic or immunotherapeutic agents; and maybe used after other regimen(s) of treatment is concluded. Examples ofchemotherapeutic agents which may be used in combination with the DRUGof the invention include but are not limited to anthracyclines,alkylating agents (e.g., mitomycin C), alkyl sulfonates, aziridines,ethylenimines, methylmelamines, nitrogen mustards, nitrosoureas,antibiotics, antimetabolites, folic acid analogs (e.g., dihydrofolatereductase inhibitors such as methotrexate), purine analogs, pyrimidineanalogs, enzymes, podophyllotoxins, platinum-containing agents,interferons, and interleukins. Particular examples of other therapeuticagents which may be used in combination with the DRUG of the inventioninclude BCR-ABL inhibitors, e.g., imatinib, nilotinib, dasatinib,dosutinib, radotinib, asciminib((R)-N-(4-(chlorodifluoromethoxy)phenyl)-6-(3-hydroxypyrrolidin-1-yl)-5-(1H-pyrazol-5-yl)nicotinamide),ponatinib and bafetinib. In an embodiment, the dose of nilotinib is10-50 mg/kg, bosutinib is 500 mg, Imatinib is 50-200 mg/kg, asciminib is90-130 mg/kg, dasatinib is 5-20 mg/kg or ponatinib is 2-10 mg/kg.BCR-ABL can contain one or more mutations. These mutations include butare not limited to V299L, T315I, F317I, F317L, Y253F, Y253H, E255K,E255V, F359C and F359V.

Particular examples of known chemotherapeutic agents which are used inan embodiment in combination with the DRUG of the invention include, butare not limited to, busulfan, improsulfan, piposulfan, benzodepa,carboquone, meturedepa, uredepa, altretamine, triethylenemelamine,triethylenephosphoramide, triethylenethiophosphoramide,trimethylolomelamine, chlorambucil, chlornaphazine, cyclophosphamide,estramustine, ifosfamide, mechlorethamine, mechlorethamine oxidehydrochloride, melphalan, novembichin, phenesterine, prednimustine,trofosfamide, uracil mustard, carmustine, chlorozotocin, fotemustine,lomustine, nimustine, ranimustine, dacarbazine, mannomustine,mitobronitol, mitolactol, pipobroman, aclacinomycins, actinomycin F(1),anthramycin, azaserine, bleomycin, cactinomycin, carubicin,carzinophilin, chromomycin, dactinomycin, daunorubicin, daunomycin,6-diazo-5-oxo-1-norleucine, doxorubicin, epirubicin, mitomycin C,mycophenolic acid, nogalamycin, olivomycin, peplomycin, plicamycin,porfiromycin, puromycin, streptonigrin, streptozocin, tubercidin,ubenimex, zinostatin, zorubicin, denopterin, methotrexate, pteropterin,trimetrexate, fludarabine, 6-mercaptopurine, thiamiprine, thioguanine,ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine,dideoxyuridine, doxifluridine, enocitabine, floxuridine, fluorouracil,tegafur, L-asparaginase, pulmozyme, aceglatone, aldophosphamideglycoside, aminolevulinic acid, amsacrine, bestrabucil, bisantrene,carboplatin, cisplatin, defofamide, demecolcine, diaziquone,elfornithine, elliptinium acetate, etoglucid, etoposide, flutamide,gallium nitrate, hydroxyurea, interferon-alpha, interferon-beta,interferon-gamma, interleukin-2, lentinan, lonidamine, prednisone,dexamethasone, leucovorin, mitoguazone, mitoxantrone, mopidamol,nitracrine, pentostatin, phenamet, pirarubicin, podophyllinic acid,2-ethylhydrazide, procarbazine, razoxane, sizofiran, spirogermanium,paclitaxel, tamoxifen, teniposide, tenuazonic acid, triaziquone,2,2′,2″-trichlorotriethylamine, urethane, vinblastine, vincristine, andvindesine.

In one embodiment, the DRUG of invention is used in combination withother antineoplastic compounds. Such compounds include, but are notlimited to ribonucleotide reductase inhibitors, topoisomerase Iinhibitors; JAK inhibitors, such as ruxolitinib; smoothened inhibitors,such as LDE225; interferon; topoisomerase II inhibitors; microtubuleactive compounds; alkylating compounds; histone deacetylase inhibitors;mTOR inhibitors, such as RAD001; antineoplastic antimetabolites; platincompounds; compounds targeting/decreasing a protein or lipid kinaseactivity methionine aminopeptidase inhibitors; biological responsemodifiers; inhibitors of Ras oncogenic isoforms; telomerase inhibitors;proteasome inhibitors; compounds used in the treatment of hematologicmalignancies, such as FLUDARABINE; compounds which target, decrease orinhibit the activity of PKC, such as midostaurin; HSP90 inhibitors suchas 17-AAG (17-allylaminogeldanamycin, NSC330507), 17-DMAG(17-dimethylaminoethylamino-17-demethoxy-geldanamycin, NSC707545),IPI-504, CNF1010, CNF2024, CNF1010 from Conforma Therapeutics, HSP990and AUY922; temozolomide (TEMODAL); kinesin spindle protein inhibitors,such as SB715992 or SB743921 from GlaxoSmithKline, orpentamidine/chlorpromazine from CombinatoRx; PI3K inhibitors, such asBEZ235, BKM120 or BYL719; MEK inhibitors such as ARRY142886 from ArrayPioPharma, AZD6244 from AstraZeneca, PD181461 from Pfizer, leucovorin,EDG binders, antileukemia compounds, Sadenosylmethionine decarboxylaseinhibitors, antiproliferative antibodies or other chemotherapeuticcompounds. Further, alternatively or in addition they may be used incombination with ionizing radiation. Further, alternatively or inaddition they may be used in combination with JAK inhibitors, such asruxolitinib. In one embodiment, the DRUG of invention is used incombination with smoothened inhibitors, such as LDE225. In oneembodiment, the DRUG of invention is used in combination withinterferon.

The DRUG of the invention may be used to treat primary, relapsed,transformed, or refractory forms of cancer, including the development ofresistance, such as mutations in BCR-ABL leading to resistance. Often,patients with relapsed cancers have undergone one or more treatmentsincluding chemotherapy, radiation therapy, bone marrow transplants,hormone therapy, surgery, and the like. Of the patients who respond tosuch treatments, they may exhibit stable disease, a partial response(i.e. the tumor or a cancer marker level diminishes by at least about50%), or a complete response (i.e. the tumor as well as markers becomeundetectable). In either of these scenarios, the cancer may subsequentlyreappear, signifying a relapse of the cancer.

In one embodiment the one or more chemotherapeutic agent is a VEGFinhibitor (e.g., an inhibitor of one or more of VEGFR (e.g., VEGFR-1,VEGFR-2, or VEGFR-3) or VEGF).

Exemplary VEGFR pathway inhibitors that can be used in combination withan IL-1β binding antibody or a functional fragment thereof, suitablygevokizumab, for use in the treatment of cancer, especially cancer withpartial inflammatory basis, e.g., CML, include, e.g., bevacizumab (alsoknown as rhuMAb VEGF or AVASTIN®), ramucirumab (Cyramza®),ziv-aflibercept (Zaltrap®), cediranib (RECENTIN™, AZD2171), lenvatinib(Lenvima®), vatalanib succinate, axitinib (INLYTA®); brivanib alaninate(BMS-582664,(S)-((R)-1-(4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-5-methylpyrrolo[2,1-f][1,2,4]triazin-6-yloxy)propan-2-yl)2-aminopropanoate);sorafenib (NEXAVAR®); pazopanib (VOTRIENT®); sunitinib malate (SUTENT®);cediranib (AZD2171, CAS 288383-20-1); vargatef (BIBF1120, CAS928326-83-4); Foretinib (GSK1363089); telatinib (BAY57-9352, CAS332012-40-5); apatinib (YN968D1, CAS 811803-05-1); imatinib (GLEEVEC®);ponatinib (AP24534, CAS 943319-70-8); tivozanib (AV951, CAS475108-18-0); regorafenib (BAY73-4506, CAS 755037-03-7); brivanib(BMS-540215, CAS 649735-46-6); vandetanib (CAPRELSA® or AZD6474);motesanib diphosphate (AMG706, CAS 857876-30-3,N-(2,3-dihydro-3,3-dimethyl-1H-indol-6-yl)-2-[(4-pyridinylmethyl)amino]-3-pyridinecarboxamide,described in PCT Publication No. WO 02/066470); semaxanib (SU5416),linfanib (ABT869, CAS 796967-16-3); cabozantinib (XL184, CAS849217-68-1); lestaurtinib (CAS 111358-88-4);N-[5-[[[5-(1,1-dimethylethyl)-2-oxazolyl]methyl]thio]-2-thiazolyl]-4-piperidinecarboxamide(BMS38703, CAS 345627-80-7);(3R,4R)-4-amino-1-((4-((3-methoxyphenyl)amino)pyrrolo[2,1-f][1,2,4]triazin-5-yl)methyl)piperidin-3-ol(BMS690514);N-(3,4-Dichloro-2-fluorophenyl)-6-methoxy-7-[[(3aα,5β,6aα)-octahydro-2-methylcyclopenta[c]pyrrol-5-yl]methoxy]-4-quinazolinamine(XL647, CAS 781613-23-8);4-methyl-3-[[1-methyl-6-(3-pyridinyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl]amino]-N-[3-(trifluoromethyl)phenyl]-benzamide(BHG712, CAS 940310-85-0); and endostatin (ENDOSTAR®).

In one embodiment the one or more chemotherapeutic agent is anti-VEGFantibody. In one embodiment the one or more chemotherapeutic agent isanti-VEGF inhibitor of small molecule weight.

In one embodiment the one or more chemotherapeutic agent is a VEGFinhibitor is selected from the list consisting of bevacizumab,ramucirumab and ziv-aflibercept. In one preferred embodiment the VEGFinhibitor is bevacizumab.

In one embodiment the one or more chemotherapeutic agent is FOLFIRI plusbevacizumab or FOLFOX plus bevacizumab or XELOX plus bevacizumab.

In one embodiment the one or more therapeutic agent, e.g., agent is acheckpoint inhibitor, preferably a PD-1 or PD-L1 inhibitor, preferablyselected from the group consisting of nivolumab, pembrolizumab,atezolizumab, avelumab, durvalumab and spartalizumab (PDR-001). In onepreferred embodiment embodiment the one or more therapeutic agent ispembrolizumab. In one preferred embodiment embodiment the one or morechemotherapeutic agent is nivolumab.

In one preferred embodiment the one or more therapeutic agent isatezolizumab. In one further preferred embodiment the one or moretherapeutic agent, e.g., chemotherapeutic agent is atezolizumab andcobimetinib.

In one preferred embodiment the one or more chemotherapeutic agent is aa tyrosine kinase inhibitor. In one embodiment said tyrosine kinaseinhibitor is an EGF pathway inhibitor, preferably an inhibitor ofEpidermal Growth Factor Receptor (EGFR). Preferably the EGFR inhibitoris chosen from one of more of erlotinib (Tarceva®), gefitinib (Iressa®),cetuximab (Erbitux®), panitumumab (Vectibix®), necitumumab (Portrazza®),dacomitinib, nimotuzumab, imgatuzumab, osimertinib (Tagrisso®),lapatinib (TYKERB®, TYVERB®). In one embodiment said EGFR inhibitor iscetuximab. In one embodiment said EGFR inhibitor is panitumumab.

In one embodiment, the EGFR inhibitor is nazartinib, i.e.(R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d]imidazol-2-yl)-2-methylisonicotinamide(Compound A40) or a compound disclosed in PCT Publication No. WO2013/184757.

The above disclosed embodiments for gevokizumab or a functional fragmentthereof are suitably applicable for canakinumab or a functional fragmentthereof.

All the disclosed uses throughout this application, including but notlimited to, doses and dosing regimens, combinations, route ofadministration and biomarkers can be applied to the treatment of CML. Inone embodiment, canakinumab is administered at a dose of from about 200mg to about 450 mg per treatment, wherein canakinumab is administeredpreferably about every 3 weeks or preferably about monthly. In oneembodiment, canakinumab is administered at a dose of 200 mg about every3 weeks or about every 4 weeks, preferably subcutaneously. In oneembodiment, canakinumab is administered at a dose of about 250 mg aboutevery 3 weeks or about every 4 weeks, preferably subcutaneously. In oneembodiment, gevokizumab is administered at a dose of from about 90 mg toabout 200 mg per treatment, wherein gevokizumab is administeredpreferably about every 3 weeks or preferably about monthly. In oneembodiment, gevokizumab is administered at a dose of about 120 mg aboutevery 3 weeks or about monthly, preferably intravenously.

In one embodiment, the present invention provides gevokizumab or afunctional fragment thereof, for use in the treatment of CML, whereingevokizumab, or a functional fragment thereof, is administered incombination with one or more therapeutic agent, e.g., chemotherapeuticagent. In one embodiment the therapeutic agent, e.g., chemotherapeuticagent is the standard of care agent for CML. In one embodiment the oneor more chemotherapeutic agent is selected from imatinib, nilotinib,dasatinib, bosutinib, radotinib, asciminib, ponatinib and bafetinib.Depending on the patient condition, at least one, at least two or atleast three chemotherapeutic agents can be selected from the list above,to be combined with gevokizumab.

In one embodiment the one or more therapeutic agent is a checkpointinhibitor, wherein preferably is a PD-1 or PD-L1 inhibitor, whereinpreferably selected from the group consisting of nivolumab,pembrolizumab, atezolizumab, avelumab, durvalumab and spartalizumab(PDR-001).

In one embodiment the one or more therapeutic agent is nivolumab. In oneembodiment the one or more chemotherapeutic agent is nivolumab plus andipilimumab. In one further embodiment said combination is used for firstor second line treatment of CML.

In one embodiment, gevokizumab or a functional fragment thereof is used,alone or preferably in combination, in first line treatment of CML. Inone embodiment gevokizumab or a functional fragment thereof is used,alone or preferably in combination, in second or third line treatment ofCML. In one embodiment, gevokizumab or a functional fragment thereof,alone or preferably in combination, is used in the treatment of CML.

The above disclosed embodiments for gevokizumab or a functional fragmentthereof are suitably applicable for canakinumab or a functional fragmentthereof.

In one aspect of this invention canakinumab or a functional fragmentthereof is administered intravenously. In one aspect of this inventioncanakinumab or a functional fragment thereof is preferably administeredsubcutaneously. Both administration routes are applicable to each andevery canakinumab related embodiments disclosed in this applicationunless in embodiments wherein the administration route is specified.

In one aspect of this invention gevokizumab or a functional fragmentthereof is administered subcutaneously. In one aspect of this inventiongevokizumab or a functional fragment thereof is preferably administeredintravenously. Both administration routes are applicable to each andevery gevokizumab related embodiments disclosed in this applicationunless in embodiments wherein the administration route is specified.

Canakinumab can be prepared as a medicament in a lyophilized form forreconstitution. In one embodiment canakinumab is provided in the form oflyophilized form for reconstitution containing at least about 200 mgdrug per vial, preferably not more than about 250 mg, preferably notmore than about 225 mg in one vial.

In one aspect the present invention provides canakinumab or gevokizumabfor use in treating and/or preventing a cancer in a patient in needthereof, comprising administering a therapeutically effective amount tothe patient, wherein the cancer has at least a partial inflammatorybasis, e.g., CML, and wherein canakinumab or gevokizumab is administeredby a prefilled syringe or by an auto-injector. Preferably the prefilledsyringe or the auto-injector contains the full amount of therapeuticallyeffective amount of the drug. Preferably the prefilled syringe or theauto-injector contains about 200 mg of canakinumab.

Efficacy and Safety

Due to its good safety profile, canakinumab or gevokizumab can beadministered to a patient for a long period of time, providing andmaintaining the benefit of suppressing IL-1β mediated inflammation.Furthermore due to its anti-cancer effect, either used in monotherapy orin combination with one or more therapeutic agents, patients life can beextended, including but not limited to extended duration of DFS, PFS,OS, hazard rate reduction, than without the Treatment of the Invention.The term “Treatment of the Invention”, as used in the this application,refers to DRUG of the invention, suitably canakinumab or gevokizumab,administered according to the dosing regimen, as taught in thisapplication. Preferably the clinical efficacy is achieved at a dose ofabout 200 mg canakinumab administered about every 3 weeks or aboutmonthly, preferably for at least about 6 months, preferably at leastabout 12 months, preferably at least about 24 months, preferably aboutup to 2 years, preferably about up to 3 years. Preferably the result isachieved at a dose of about 30 mg-120 mg gevokizumab administered aboutevery 3 weeks or about monthly, preferably for at least about 6 months,preferably at least about 12 months, preferably at least about 24months, preferably about up to 2 years, preferably about up to 3 years.In one embodiment Treatment of the Invention is the sole treatment. Inone embodiment Treatment of the Invention is added on top of the SoCtreatment for the cancer indication. While the SoC treatment evolveswith time, the SoC treatment as used here should be understood as notincluding DRUG of the invention.

Thus in one aspect the present invention provides an IL-1β bindingantibody or functional fragment thereof, suitably canakinumab orgevokizumab, for use in the treatment and/or prevention of cancer, e.g.,cancer that has at least a partial inflammatory basis, e.g., CML, in apatient, wherein a therapeutically effective amount of an IL-1β bindingantibody or a functional fragment thereof is administered in the patientfor at least about 6 months, preferably at least about 12 months,preferably at least about 24 months. In one embodiment the cancerexcludes lung cancer, especially excludes NSCLC, especially excludespost-surgery NSCLC, in which the cancer has been resect, suitably notlonger than about 2 months, preferably not longer than about one month.

In one aspect, the present invention provides an IL-1β binding antibodyor functional fragment thereof, suitably canakinumab or gevokizumab, foruse in the treatment of cancer, e.g., cancer that has at least a partialinflammatory basis, e.g., CML, in a patient, wherein the hazard rate ofcancer mortality of the patient is reduced by at least about 10%, atleast about 20%, at least about 30%, at least about 40% or at leastabout 50%, preferably compared to not receiving Treatment of theInvention.

The term “not receiving Treatment of the Invention”, as used throughoutthe application, include patient did not receive any drug at all andpatient received only treatment, considered as SoC at the time, withoutthe DRUG of the invention. As a skilled person would understand, theclinical efficacy is typically not tested within the same patient,receiving or not receiving the Treatment of the Invention, rather testedin clinical trial settings with treatment group and placebo group.

In one embodiment the overall survival (OS, defined as the time from thedate of randomization to the date of death due to any cause) in thepatient is at least about one month, at least about 3 months, at leastabout 6 months, at least about 12 months longer than not receivingTreatment of the Invention. In one embodiment the OS is at least about12 months, preferably at least about 24 months, longer in the adjuvanttreatment setting. In one embodiment the OS is at least 4 months,preferably at least about 6 months, at least about 12 months longer inthe first line treatment setting. In one embodiment the OS is at leastabout one month, at least about 3 months, preferably at least about 6months longer in the 2^(nd)/3^(rd) line treatment setting.

In one embodiment the overall survival in the patient receivingTreatment of the Invention is at least about 2 years, at least about 3years, at least about 5 years, at least about 8 years, at least about 10years in the adjuvant treatment setting. In one embodiment the overallsurvival in the patient receiving Treatment of the Invention is at leastabout 6 month, at least about one year, at least about 3 years in thefirst line treatment setting. In one embodiment the overall survival inthe patient receiving Treatment of the Invention is at least about 3month, at least about 6 months, at least about one year in the2^(nd)/3^(rd) line treatment setting.

In one embodiment the progression free survival (PFS) period of thepatient receiving Treatment of the Invention is extended by at leastabout one months, at least about 2 months, at least about 3 months, atleast about 6 months, at least about 12 months, preferably compared tonot receiving Treatment of the Invention. In one embodiment PFS isextended by at least about 6 months, preferably at least about 12 monthsin the first line treatment settings. In one embodiment PFS is extendedby at least about one month, at least about 3 months, at least about 6months in the second line treatment settings.

In one embodiment the patient receiving Treatment of the Invention hasat least about 3 months, at least about 6 months, at least about 12months, or at least about 24 months progression free survival.

Normally clinical efficacy, including but not limited to DFS, PFS, HRreduction, OS, can be demonstrated in clinical trials comparingtreatment group and placebo group. In the placebo group patients receiveno drug at all or receive SoC treatment. In the treatment group patientsreceive DRUG of the invention either as monotherapy or added to the SoCtreatment. Alternatively in the placebo group patients receive SoCtreatment and in the treatment group patients receive DRUG of theinvention.

Even though the clinical outcome, such as duration of DFS or the HRreduction of cancer mortality, is described as a number based onstatistical analysis of a clinical trial, one of ordinary skill wouldreadily extrapolate these statistics to treatments for an individualpatient, as claimed, since it is expected the DRUG of the inventionwould achieve a similar clinical outcome in a portion of the individualpatients receiving Treatment of the Invention, for example in 95% of thepatients, when clinical trials have demonstrated statisticalsignificance (p_≤0.05)); or for example in 50% of the patients, whenclinical trials have provided mean value, such as mean PFS is 24 months.

IL-1β blockade could affect a patients' immune system in combatinginfection. Thus in one aspect the present invention provides an IL-1βbinding antibody or a functional fragment thereof, suitably canakinumabor gevokizumab, for use in the treatment and/or prevention of cancer,e.g., cancer having at least a partial inflammatory basis, e.g., CML,and wherein the patient is not at high risk of developing serious ainfection due to the Treatment of the Invention. Patients would be athigh risk of developing serious infection due to the Treatment of theInvention in the following, but not limited to, the followingsituations: (a) Patients have an active infection requiring medicalintervention. The term “active infection requiring medical intervention”is understood as the patient is currently taking or have been taking orhave just finished taken for less than about one month or less thanabout two weeks, any anti-viral and/or any anti-bacterial medicines; (b)Patients have latent tuberculosis and/or a history of tuberculosis.

To manage the inhibition of the immune system by IL-1β blockade, it iscautioned that the IL-1β binding antibody or a functional fragmentthereof is not administered concomitantly with a TNF inhibitor.Preferably a TNF inhibitor is selected from a group consisting ofEnbrel® (etanercept), Humira® (adalimumab), Remicade® (infliximab),Simponi® (golimumab), and Cimzia® (certolizumab pegol). It is alsocautioned that the IL-1β binding antibody or a functional fragmentthereof is not administered concomitantly with another IL-1 blocker,wherein preferably said IL-1 blocker is selected from a group consistingof Kineret® (anakinra) and Arcalyst® (rilonacept). Furthermore it isonly one IL-1β binding antibody or a functional fragment thereof isadministered in the treatment/prevention of cancer. For examplecanakinumab is not administered in combination with gevokizumab.

When canakinumab is administered into patients, it is likely that somepatients will develop anti-canakinumab antibody (anti-drug antibody,ADA), which needs to be monitored for safety and efficacy reasons. Inone aspect the present invention provides canakinumab for use in thetreatment and/or preventing cancer, e.g., cancer having at least apartial inflammatory basis, e.g., CML, wherein the chance of the patientdeveloping ADA is less than about 1%, less than about 0.7%, less thanabout 0.5%, or less than about 0.4%. In one embodiment the antibody isdetected by the method as described in Example 10. In one embodiment theantibody is detection is performed at about 3 months, at about 6 monthsor at about 12 month from the first administration of canakinumab.

Canakinumab can be administered in a reconstituted formulationcomprising canakinumab at a concentration of about 50-200 mg/ml, about50-300 mM sucrose, about 10-50 mM histidine, and about 0.01-0.1%surfactant and wherein the pH of the formulation is about 5.5-7.0.Canakinumab can be administered in a reconstituted formulationcomprising canakinumab at a concentration of about 50-200 mg/ml, about270 mM sucrose, about 30 mM histidine and about 0.06% polysorbate 20 or80, wherein the pH of the formulation is about 6.5.

Canakinumab can also be administered in a liquid formulation comprisingcanakinumab at a concentration of about 50-200 mg/ml, a buffer systemselected from the group consisting of citrate, histidine and sodiumsuccinate, a stabilizer selected from the group consisting of sucrose,mannitol, sorbitol, arginine hydrochloride, and a surfactant and whereinthe pH of the formulation is about 5.5-7.0. Canakinumab can also beadministered in a liquid formulation comprising canakinumab at aconcentration of about 50-200 mg/ml, about 50-300 mM mannitol, about10-50 mM histidine and about 0.01-0.1% surfactant, and wherein the pH ofthe formulation is about 5.5-7.0. Canakinumab can also be administeredin a liquid formulation comprising canakinumab at a concentration ofabout 50-200 mg/ml, about 270 mM mannitol, about 20 mM histidine andabout 0.04% polysorbate 20 or 80, wherein the pH of the formulation isabout 6.5.

When administered subcutaneously, canakinumab can be administered to thepatient in a liquid form contained in a prefilled syringe or as alyophilized form for reconstitution.

In one aspect, the present invention provides high sensitivityC-reactive protein (hsCRP) for use as a biomarker in the treatmentand/or prevention of cancer, e.g., cancer having at least a partialinflammatory basis, including but not limited to CML, with an IL-1βinhibitor, e.g., IL-1β binding antibody or a functional fragmentthereof. Typically cancers that have at least a partial inflammatorybasis include but are not limited to lung cancer, especially NSCLC, CML,colorectal cancer, melanoma, gastric cancer (including esophagealcancer), renal cell carcinoma (RCC), breast cancer, hepatocellularcarcinoma (HCC), prostate cancer, bladder cancer, AML, multiple myelomaand pancreatic cancer. Consistent with prior work indicating a stronginflammatory component to certain cancers, hsCRP levels in the CANTOStrial population were elevated at baseline among those who werediagnosed with lung cancer during follow-up compared to those whoremained free of any cancer diagnosis (6.0 versus 4.2 mg/L, P<0.001).Thus the level of hsCRP is possibly relevant in determining whether apatient with diagnosed lung cancer, undiagnosed lung cancer or is atrisk of developing lung cancer should be treated with an IL-1βinhibitor, IL-1β binding antibody or a functional fragment thereof. In apreferred embodiment, said IL-1β binding antibody or a fragment thereofis canakinumab or a fragment thereof or gevokizumab or a fragmentthereof. Similarly the level of hsCRP is possibly relevant indetermining whether a patient with cancer having at least a partialinflammatory basis, diagnosed or undiagnosed, should be treated with anIL-1β inhibitor, IL-1β binding antibody or a functional fragmentthereof. In a preferred embodiment, said IL-1β binding antibody iscanakinumab or gevokizumab.

Thus the present invention provides high sensitivity C-reactive protein(hsCRP) for use as a biomarker in the treatment and/or prevention ofcancer having at least a partial inflammatory basis, including CML, in apatient with an IL-1β inhibitor, IL-1β binding antibody or a functionalfragment thereof, wherein said patient is eligible for the treatmentand/or prevention if the level of high sensitivity C-reactive protein(hsCRP) is equal to or higher than about 2 mg/L, or equal to or higherthan about 3 mg/L, or equal to or higher than about 4 mg/L, or equal toor higher than about 5 mg/L, or equal to or higher than about 6 mg/L,equal to or higher than about 7 mg/L, equal to or higher than about 8mg/L, equal to or higher than about 9 mg/L, or equal to or higher thanabout 10 mg/L, equal to or higher than about 12 mg/L, equal to or higherthan about 15 mg/L, equal to or higher than about 20 mg/L or equal to orhigher than about 25 mg/L as assessed prior to the administration of theIL-1β binding antibody or a functional fragment thereof. In a preferredembodiment, said patient has hsCRP level equal to or higher than about 4mg/L. In a preferred embodiment, said patient has hsCRP level equal toor higher than about 6 mg/L. In a preferred embodiment, said patient hashsCRP level equal to or higher than about 10 mg/L.

In analyses of combined canakinumab doses, compared to placebo, theobserved hazard ratio for lung cancer among those who achieved hsCRPreductions greater than the median value of 1.8 mg/L at 3 months was0.29 (95% CI 0.17-0.51, P<0.0001), better than the effect observed forthose who achieved hsCRP reductions less than the median value (HR 0.83,95% CI 0.56-1.22, P=0.34).

Thus in one aspect, the present invention relates to the use of thedegree of reduction of the hsCRP as a prognostic biomarker to guidephysician in continuing or discontinuing with the treatment of an IL-1βinhibitor, an IL-1β binding antibody or a functional fragment thereof,especially canakinumab or gevokizumab. In one embodiment, the presentinvention provides the use of an IL-1β inhibitor, an IL-1β bindingantibody or a functional fragment thereof, in the treatment and/orprevention of cancer having at least a partial inflammatory basis,including CML, wherein such treatment or prevention is continued whenthe level of hsCRP is reduced by at least about 0.8 mg/L, at least about1 mg/L, at least about 1.2 mg/L, at least about 1.4 mg/L, at least about1.6 mg/L, at least about 1.8 mg/L, at least about 3 mg/L or at leastabout 4 mg/L, at least about 3 months, preferably about 3 months afterfirst administration of the IL-1β binding antibody or functionalfragment thereof. In one embodiment, the present invention provides theuse of an IL-1β inhibitor, IL-1β binding antibody or a functionalfragment thereof, in the treatment and/or prevention of cancer having atleast a partial inflammatory basis, including lung cancer, wherein suchtreatment or prevention is discontinued when the level of hsCRP isreduced by less than about 0.8 mg/L, less than about 1 mg/L, less thanabout 1.2 mg/L, less than about 1.4 mg/L, less than about 1.6 mg/L, lessthan about 1.8 mg/L at about 3 months from the beginning of thetreatment at an appropriate dosing with the IL-1β binding antibody orfunctional fragment thereof. In a further embodiment the appropriatedosing of canakinumab is about 50 mg, about 150 mg or about 300 mg,which is administered about every 3 months. In a further embodiment theappropriate dosing of canakinumab is about 300 mg administered twiceover a two-week period and then about every three months. In oneembodiment, the IL-1β binding antibody or a functional fragment thereofis canakinumab or a functional fragment thereof, wherein saidcanakinumab is administered at a dose of about 200 mg about every 3weeks or about 200 mg about monthly. In one embodiment, the IL-1βbinding antibody or a functional fragment thereof is gevokizumab or afunctional fragment thereof, wherein said gevokizumab is administered ata dose of about 60 mg to about 90 mg or about 120 mg about every 3 weeksor about monthly.

In one aspect, the present invention provides the use of the reducedhsCRP level as a prognostic biomarker to guide a physician in continuingor discontinuing with the treatment of an IL-1β binding antibody or afunctional fragment thereof, especially canakinumab or gevokizumab. Inone embodiment, such treatment and/or prevention with the IL-1β bindingantibody or a functional fragment thereof is continued when the level ofhsCRP is reduced below about 10 mg/L, reduced below about 8 mg/L,reduced below about 5 mg/L, reduced below about 3.5 mg/L, below about 3mg/L, below about 2.3 mg/L, below about 2 mg/L or below about 1.8 mg/Lassessed at least about 3 months from first administration of the IL-1βbinding antibody or a functional fragment thereof. In one embodiment,such treatment and/or prevention with the IL-1β binding antibody or afunctional fragment thereof is discontinued when the level of hsCRP isnot reduced below about 3.5 mg/ml, below about 3 mg/L, below about 2.3mg/L, below about 2 mg/L or below about 1.8 mg/L assessed at least about3 months from first administration of the IL-1β binding antibody or afunctional fragment thereof. In a further embodiment the appropriatedosing is canakinumab at about 300 mg administered twice over a two-weekperiod and then about every three months. In one embodiment, the IL-1βbinding antibody or a functional fragment thereof is canakinumab or afunctional fragment thereof, wherein said canakinumab is administered ata dose of about 200 mg about every 3 weeks or about 200 mg about monthlyor about 300 mg about monthly. In one embodiment, the IL-1β bindingantibody or a functional fragment thereof is gevokizumab or a functionalfragment thereof, wherein said gevokizumab is administered at a dose ofabout 60 mg to about 90 mg or about 120 mg about every 3 weeks or aboutmonthly.

Without wishing to be being bound by theory, it is hypothesized that theinhibition of IL-1β pathway can lead to inhibition or reduction of tumormetastasis. Until now there have been no reports on the effects ofcanakinumab on metastasis. Data presented in example 3 demonstrate thatIL-1β activates different pro-metastatic mechanisms at the primary sitecompared with the metastatic site: Endogenous production of IL-1β bybreast cancer cells promotes epithelial to mesenchymal transition (EMT),invasion, migration and organ specific homing. Once tumor cells arrivein the bone environment contact between tumor cells and osteoblasts orbone marrow cells increase IL-1β secretion from all three cell types.These high concentrations of IL-1β cause proliferation of the bonemetastatic niche by stimulating growth of disseminated tumor cells intoovert metastases. These pro-metastatic processes are inhibited byadministration of anti-IL-1β treatments, such as canakinumab.

Therefore, targeting IL-1β with an IL-1β binding antibody represents anovel therapeutic approach for cancer patients at risk of progressing tometastasis by preventing seeding of new metastases from establishedtumors and retaining tumor cells already disseminated in the bone in astate of dormancy. The models described have been designed toinvestigate bone metastasis and although the data show a strong linkbetween IL-1β expression and bone homing, it does not exclude IL-1βinvolvement in metastasis to other sites.

Accordingly, in one aspect, the present invention provides an IL-1βbinding antibody or a functional fragment thereof for use in a patientin need thereof in the treatment of a cancer having at least partialinflammatory basis, wherein said IL-1β binding antibody or a functionalfragment thereof is administered at a dose sufficient to inhibitmetastasis in said patient. Typically cancer having at least partialinflammatory basis includes but is not limited to CML.

In one embodiment said dose sufficient to inhibit metastasis comprisesan IL-1β binding antibody or a functional fragment thereof to beadministered in the range of about 30 mg to about 750 mg per treatment,alternatively about 100 mg-600 mg, about 100 mg to about 450 mg, about100 mg to about 300 mg, alternatively about 150 mg-about 600 mg, about150 mg to about 450 mg, about 150 mg to about 300 mg, preferably about150 mg to about 300 mg; alternatively at least about 150 mg, at leastabout 180 mg, at least about 250 mg, at least about 300 mg pertreatment. In one embodiment the patient with a cancer that has at leasta partial inflammatory basis, including lung cancer, receives eachtreatment about every 2 weeks, about every three weeks, about every fourweeks (monthly), about every 6 weeks, about bimonthly (every 2 months)or about quarterly (every 3 months). In one embodiment the range of DRUGof the invention is about 90 mg to about 450 mg. In one embodiment saidDRUG of the invention is administered about monthly. In one embodimentsaid DRUG of the invention is administered about every 3 weeks.

In one embodiment the IL-1β binding antibody is canakinumab administeredat a dose sufficient to inhibit metastasis, wherein said dose is in therange of about 100 mg to about 750 mg per treatment, alternatively about100 mg-600 mg, about 100 mg to about 450 mg, about 100 mg to about 300mg, alternatively about 150 mg-600 mg, about 150 mg to about 450 mg,about 150 mg to about 300 mg, alternatively at least about 150 mg, atleast about 200 mg, at least about 250 mg, at least about 300 mg pertreatment. In one embodiment the patient with cancer having at least apartial inflammatory basis, including lung cancer, receives eachtreatment about every 2 weeks, about every 3 weeks, about every 4 weeks(monthly), about every 6 weeks, about bimonthly (every 2 months) orabout quarterly (every 3 months). In one embodiment the patient withcancer receives canakinumab about monthly. In one embodiment thepreferred dose range of canakinumab is about 200 mg to about 450 mg,further preferred about 300 mg to about 450 mg, further preferred about350 mg to about 450 mg. In one embodiment the preferred dose range ofcanakinumab is about 200 mg to about 450 mg about every 3 weeks or aboutmonthly. In one embodiment the preferred dose of canakinumab is about200 mg about every 3 weeks. In one embodiment the preferred dose ofcanakinumab is about 200 mg about monthly. In one embodiment canakinumabis administered subcutaneously or intravenously, preferablysubcutaneously.

In one embodiment, the IL-1β binding antibody is gevokizumabadministered at a dose sufficient to inhibit metastasis, wherein saiddose is in the range of about 30 mg to about 450 mg per treatment,alternatively about 90 mg-450 mg, about 90 mg to about 360 mg, about 90mg to about 270 mg, about 90 mg to about 180 mg; alternatively about 120mg-450 mg, about 120 mg to about 360 mg, about 120 mg to about 270 mg,about 120 mg to about 180 mg, alternatively about 150 mg-450 mg, about150 mg to about 360 mg, about 150 mg to about 270 mg, about 150 mg toabout 180 mg; alternatively about 180 mg-450 mg, about 180 mg to about360 mg, about 180 mg to about 270 mg; alternatively at least about 150mg, at least about 180 mg, at least about 240 mg, at least about 270 mgper treatment. In one embodiment the patient with cancer that has atleast a partial inflammatory basis, including lung cancer, receivestreatment about every 2 weeks, about every 3 weeks, about monthly, aboutevery 6 weeks, about bimonthly (every 2 months) or about quarterly(every 3 months). In one embodiment the patient with cancer that has atleast a partial inflammatory basis, including lung cancer, receives atleast one, preferably one treatment per month. In one embodiment thepreferred range of gevokizumab is about 150 mg to about 270 mg. In oneembodiment the preferred range of gevokizumab is about 60 mg to about180 mg, further preferred about 60 mg to about 90 mg. In one embodimentthe preferred schedule is about every 3 weeks. In one embodiment thepreferred schedule is about monthly. In one embodiment the patientreceives gevokizumab about 60 mg to about 90 mg about every 3 weeks. Inone embodiment the patient receives gevokizumab about 60 mg to about 90mg about monthly. In one embodiment the patient with cancer that has atleast a partial inflammatory basis receives gevokizumab about 90 mg toabout 360 mg, about 90 mg to about 270 mg, about 120 mg to about 270 mg,about 90 mg to about 180 mg, about 120 mg to about 180 mg, about 120 mgor about 90 mg about every 3 weeks. In one embodiment the patient withcancer that has at least a partial inflammatory basis receivesgevokizumab about 90 mg to about 360 mg, about 90 mg to about 270 mg,about 120 mg to about 270 mg, about 90 mg to about 180 mg, about 120 mgto about 180 mg, about 120 mg or about 90 mg monthly. In one embodimentthe patient receives gevokizumab about 90 mg, every about 180 mg, about190 mg or about 200 mg about every 3 weeks. In one embodiment thepatient receives gevokizumab about 90 mg, about 180 mg, about 190 mg orabout 200 mg about monthly. In one embodiment the patient receivesgevokizumab about 120 mg about monthly or about every 3 weeks. In oneembodiment gevokizumab is administered subcutaneously or intravenously,preferably intravenously.

In one aspect the present invention provides an IL-1β binding antibodyor a functional fragment thereof, preferably gevokizumab or a functionalfragment thereof or canakinumab or a functional fragment thereof, foruse in the treatment of cancer in a patient, wherein the hsCRP level hasreduced to at least about 30%, preferably at least about 40%, preferablyat least about 50% compared to the baseline level (prior to treatment),or reduced to below about 10 mg/L, below about 7 mg/L, or below about 5mg/L, at about 6 months or at about 3 months or about one month afterthe first administration of the DRUG of the invention. Preferablycanakinumab or a functional fragment thereof is administered about200-450 mg, preferably about 200 mg about every 3 weeks or about every 4weeks, preferably subcutaneously. Preferably gevokizumab or a functionalfragment thereof is administered about 30-120 mg, preferably about 60-90mg about every 3 weeks or about every 4 weeks, preferably intravenously.

All the disclosed uses throughout this application, including but notlimited to, doses and dosing regimens, combinations, route ofadministration and biomarkers can be applied to the embodiment ofmetastasis inhibition. In one preferred embodiment IL-1β antibody or afunctional fragment thereof is used in combination of one or morechemotherapeutic agents, wherein said agent is an anti-Wnt inhibitor,preferably Vantictumab.

IL-1β is known to drive the induction of gene expression of a variety ofpro-inflammatory cytokines, such as IL-6 and TNF-α. In the CANTOS trial,it was observed that administration of canakinumab was associated withdose-dependent reductions in IL-6 of 25 to 43 percent (allP-values<0.0001). The present application therefore provides an IL-6inhibitor for use in the treatment and/or prevention of cancer having atleast a partial inflammatory basis, including but not limited to CML. Insome embodiments, the IL-6 inhibitor is selected from the groupconsisting of: anti-sense oligonucleotides against IL-6, IL-6 antibodiessuch as siltuximab (Sylvant®), sirukumab, clazakizumab, olokizumab,elsilimomab, gerilimzumab, WBP216 (also known as MEDI 5117), or afragment thereof, EBI-031 (Eleven Biotherapeutics), FB-704A (FountainBioPharma Inc), OP-R003 (Vaccinex Inc), IG61, BE-8, PPV-06 (Peptinov),SBP002 (Solbec), Trabectedin (Yondelis®), C326/AMG-220, olamkicept, PGE1and its derivatives, PGI2 and its derivatives, and cyclophosphamide.Another embodiment of the present invention provides an IL-6 receptor(IL-6R) (CD126) inhibitor for use in the treatment and/or prevention ofcancer having at least a partial inflammatory basis, including CML. Insome embodiments, the IL-6R inhibitor is selected from the groupconsisting of: anti-sense oligonucleotides against IL-6R, tocilizumab(Actemra®), sarilumab (Kevzara®), vobarilizumab, PM1, AUK12-20, AUK64-7,AUK146-15, MRA, satralizumab, SL-1026 (SomaLogic), LTA-001 (CommonPharma), BCD-089 (Biocad Ltd), APX007 (Apexigen/Epitomics), TZLS-501(Novimmune), LMT-28, anti-IL-6R antibodies disclosed in WO2007143168 andWO2012118813, Madindoline A, Madindoline B, and AB-227-NA.

In one aspect the present application provides an IL-6 inhibitor incombination with one or more chemotherapeutic agent for use in thetreatment of cancer having at least a partial inflammatory basis. In oneembodiment the one or more chemotherapeutic agent is a check pointinhibitor. In one embodiment said check point inhibitor is a PD-1 orPD-L1 inhibitor preferably selected from the group consisting ofnivolumab, pembrolizumab, atezolizumab, durvalumab, avelumab andspartalizumab (PDR-001).

In one embodiment the one or more chemotherapeutic agent is the standardof care chemotherapy for a defined cancer having at least a partialinflammatory basis, e.g., CML.

Other features, objects, and advantages of the invention will beapparent from the description and drawings, and from the claims.

The following Examples illustrate the invention described above; theyare not, however, intended to limit the scope of the invention in anyway.

In one embodiment, the present invention provides an IL-1β bindingantibody or a functional fragment thereof, suitably canakinumab, for usein the treatment of cancer that has at least a partial inflammatorybasis, including CML, wherein the risk for cancer that has at least apartial inflammatory basis, including CML, is reduced by at least about30%, at least about 40%, at least about 50% at about 3 months from thefirst administration compared to patient not receiving the treatment. Inone preferred embodiment, the dose of the first administration is atabout 300 mg. In one further preferred embodiment, the dose of the firstadministration is at about 300 mg followed by a second dose of about 300mg within a two-week period. Preferably the result is achieved with adose of about 200 mg canakinumab administered about every 3 weeks.Preferably the result is achieved with a dose of about 200 mgcanakinumab administered about every month.

In one embodiment, the present invention provides an IL-1β bindingantibody or functional fragment thereof, suitably canakinumab, for usein the treatment of cancer that has at least a partial inflammatorybasis, including CML, wherein the risk for CML mortality is reduced byat least about 30%, at least about 40% or at least about 50% compared toa patient not receiving the treatment. Preferably the results isachieved at a dose of about 200 mg canakinumab administered about every3 weeks or about 300 mg canakinumab administered about monthly,preferably for at least for about one year, preferably up to about 3years.

In one embodiment, the present invention provides an IL-1β bindingantibody or functional fragment thereof, suitably canakinumab or afunctional fragment thereof, suitably gevokizumab or a functionalfragment thereof for use in the treatment of cancer that has at least apartial inflammatory basis, wherein the risk for said cancer mortalityis reduced by at least about 30%, at least about 40% or at least about50% compared to a patient not receiving the treatment. Preferably theresults is achieved at a dose of about 200 mg canakinumab administeredabout every 3 weeks or about monthly, preferably for at least for aboutone year, preferably up to about 3 years. Preferably the results isachieved at a dose of about 120 mg gevokizumab administered about every3 weeks or about monthly, preferably for about at least for one year,preferably up to about 3 years. Preferably the results is achieved at adose of about 90 mg gevokizumab administered about every 3 weeks orabout monthly, preferably for at least about one year, preferably up toabout 3 years.

EXAMPLES

The Example below is set forth to aid in the understanding of theinvention but is not intended, and should not be construed, to limit itsscope in any way.

Example 1 Tumor-Derived IL-1β Induces Differential Tumor PromotingMechanisms in Metastasis Materials and Methods Cell Culture

Human breast cancer MDA-MB-231-Luc2-TdTomato (Calliper Life Sciences,Manchester UK), MDA-MB-231 (parental) MCF7, T47D (European Collection ofAuthenticated Cell Cultures (ECACC)), MDA-MB-231-IV (Nutter et al.,2014) as well as bone marrow HS5 (ECACC) and human primary osteoblastsOB1 were cultured in DMEM+10% FCS (Gibco, Invitrogen, Paisley, UK). Allcell lines were cultured in a humidified incubator under 5% C02 and usedat low passage >20.

Transfection of Tumor Cells

Human MDA-MB-231, MCF 7 and T47D cells were stably transfected tooverexpress genes IL1B or IL1R1 using plasmid DNA purified fromcompetent E. coli that have been transduced with an ORF plasmidcontaining human IL1B or IL1R1 (Accession numbers NM_000576 andNM_0008777.2, respectively) with a C-terminal GFP tag (OriGeneTechnologies Inc. Rockville Md.). Plasmid DNA purification was performedusing a PureLink™ HiPure Plasmid Miniprep Kit (ThermoFisher) and DNAquantified by UV spectroscopy before being introduced into human cellswith the aid of Lipofectamine II (ThermoFisher). Control cells weretransfected with DNA isolated from the same plasmid without IL-1B orIL-1R1 encoding sequences.

In Vitro Studies

In vitro studies were carried out with and without addition of 0-5 ng/mlrecombinant IL-1β (R&D systems, Wiesbaden, Germany) +/−50 μM IL-1Ra(Amgen, Cambridge, UK).

Cells were transferred into fresh media with 10% or 1% FCS. Cellproliferation was monitored every 24 h for up to 120 h by manual cellcounting using a 1/400 mm² hemocytometer (Hawkley, Lancing UK) or over a72 h period using an Xcelligence RTCA DP Instrument (Acea Biosciences,Inc). Tumor cell invasion was assessed using 6 mm transwell plates withan 8 μm pore size (Corning Inc) with or without basement membrane (20%Matrigel; Invitrogen).

Tumor cells were seeded into the inner chamber at a density of 2.5×10⁵for parental as well as MDA-MB-231 derivatives and 5×10⁵ for T47D inDMEM+1% FCS and 5×10⁵ OB1 osteoblast cells supplemented with 5% FCS wereadded to the outer chamber. Cells were removed from the top surface ofthe membrane 24 h and 48 h after seeding and cells that had invadedthrough the pores were stained with hematoxylin and eosin (H&E) beforebeing imaged on a Leica DM7900 light microscope and manually counted.

Migration of cells was investigated by analyzing wound closure: Cellswere seeded onto 0.2% gelatine in 6-well tissue culture plates (Costar;Corning, Inc) and, once confluent, 10 μg/ml mitomycin C was added toinhibit cell proliferation and a 50 μm scratch made across themonolayer. The percentage of wound closure was measured at 24 h and 48 husing a CTR7000 inverted microscope and LAS-AF v2.1.1 software (LeicaApplications Suite; Leica Microsystems, Wetzlar, Germany). Allproliferation, invasion and migration experiments were repeated usingthe Xcelligence RTCA DP instrument and RCTA Software (Acea Biosystems,Inc).

For co-culture studies with human bone 5×10⁵ MDA-MB-231 or T47D cellswere seeded onto tissue culture plastic or into 0.5 cm³ human bone discsfor 24 h. Media was removed and analysed for concentration of IL-1β byELISA. For co-culture with HS5 or OB1 cells, 1×10⁵ MDA-MB-231 or T47Dcells were cultured onto plastic along with 2×10⁵ HS5 or OB1 cells.Cells were sorted by FACS 24 h later and counted and lysed for analysisof IL-1β concentration. Cells were collected, sorted and counted every24 h for 120 h.

Animals

Experiments using human bone grafts were carried out in 10-week oldfemale NOD SCID mice. In IL-1β/IL-1R1 overexpression bone homingexperiments 6 to 8-week old female BALB/c nude mice were used. Toinvestigate effects of IL-1β on the bone microenvironment 10-week oldfemale C57BL/6 mice (Charles River, Kent, UK) or IL-1R1^(−/−) mice(Abdulaal et al., 2016) were used. Mice were maintained on a 12 h:12 hlight/dark cycle with free access to food and water. Experiments werecarried out with UK home office approval under project licence 40/3531,University of Sheffield, UK.

Patient Consent and Preparation of Bone Discs

All patients provided written, informed consent prior to participationin this study. Human bone samples were collected under HTA licence12182, Sheffield Musculoskeletal Biobank, University of Sheffield, UK.Trabecular bone cores were prepared from the femoral heads of femalepatients undergoing hip replacement surgery using an Isomat 4000Precision saw (Buehler) with Precision diamond wafering blade (Buehler).5 mm diameter discs were subsequently cut using a bone trephine beforestoring in sterile PBS at ambient temperature.

In Vivo Studies

To model human breast cancer metastasis to human bone implants two humanbone discs were implanted subcutaneously into 10-week old female NODSCID mice (n=10/group) under isofluorane anaesthetic. Mice received aninjection of 0.003 mg vetergesic and Septrin was added to the drinkingwater for 1 week following bone implantation. Mice were left for 4 weeksbefore injecting 1×10⁵ MDA-MB-231 Luc2-TdTomato, MCF7 Luc2 or T47D Luc2cells in 20% Martigel/79% PBS/1% toluene blue into the two hind mammaryfat pads. Primary tumor growth and development of metastases wasmonitored weekly using an IVIS (Luminol) system (Caliper Life Sciences)following sub-cutaneous injection of 30 mg/ml D-luciferin (Invitrogen).On termination of experiments mammary tumors, circulating tumor cells,serum and bone metastases were resected. RNA was processed fordownstream analysis by real time PCR, and cell lysates were taken forprotein analysis and whole tissue for histology as previously described(Nutter et al., 2014; Ottewell et al., 2014a).

For therapeutic studies in NOD SCID mice, placebo (control), 1 mg/kgIL-1Ra (anakinra®) daily or 10 mg/kg canakinumab subcutaneously every 14days were administered starting 7 days after injection of tumor cells.In BALB/c mice and C57BL/6 mice 1 mg/kg IL-1Ra was administered dailyfor 21 or 31 days or 10 mg/kg canakinumab was administered as a singlesubcutaneous injection. Tumor cells, serum, and bone were subsequentlyresected for downstream analysis.

Bone metastases were investigated following injection of 5×10⁵MDA-MB-231 GFP (control), MDA-MB-231-IV, MDA-MB-231-IL-1B-positive orMDA-MB-231-IL-1R1-positive cells into the lateral tail vein of 6 to8-week old female BALB/c nude mice (n=12/group). Tumor growth in bonesand lungs was monitored weekly by GFP imaging in live animals. Mice wereculled 28 days after tumor cell injection at which timepoint hind limbs,lungs and serum were resected and processed for microcomputed tomographyimaging (μCT), histology and ELISA analysis of bone turnover markers andcirculating cytokines as described (Holen et al., 2016).

Isolation of Circulating Tumor Cells

Whole blood was centrifuged at 10,000×g for 5 minutes and the serumremoved for ELISA assays. The cell pellet was re-suspended in 5 ml ofFSM lysis solution (Sigma-Aldrich, Pool, UK) to lyse red blood cells.Remaining cells were re-pelleted, washed 3× in PBS and re-suspended in asolution of PBS/10% FCS. Samples from 10 mice per group were pooledprior to isolation of TdTomato positive tumor cells using a MoFlow Highperformance cell sorter (Beckman Coulter, Cambridge UK) with the 470 nMlaser line from a Coherent I-90C tenable argon ion (Coherent, SantaClara, Calif.). TdTomato fluorescence was detected by a 555LP dichroiclong pass and a 580/30 nm band pass filter. Acquisition and analysis ofcells was performed using Summit 4.3 software. Following sorting cellswere immediately placed in RNA protect cell reagent (Ambion, Paisley,Renfrew, UK) and stored at −80° C. before RNA extraction. For countingnumbers of circulating tumor cells, TdTomato fluorescence was detectedusing a 561 nm laser and an YL1-A filter (585/16 emission filter).Acquisition and analysis of cells was performed using Attune N×Tsoftware.

Microcomputed Tomography Imaging

Microcomputed tomography (μCT) analysis was carried out using a Skyscan1172 x-ray-computed μCT scanner (Skyscan, Aartselar, Belgium) equippedwith an x-ray tube (voltage, 49 kV; current, 200 uA) and a 0.5-mmaluminium filter. Pixel size was set to 5.86 μm and scanning initiatedfrom the top of the proximal tibia as previously described (Ottewell etal., 2008a; Ottewell et al., 2008b).

Bone Histology and Measurement of Tumor Volume

Bone tumor areas were measured on three non-serial, H&E stained, 5 μmhistological sections of decalcified tibiae per mouse using a Leica RMRBupright microscope and Osteomeasure software (Osteometrics, Inc.Decauter, USA) and a computerised image analysis system as previouslydescribed (Ottewell et al., 2008a).

Western Blotting

Protein was extracted using a mammalian cell lysis kit (Sigma-Aldrich,Poole, UK). 30 μg of protein was run on 4-15% precast polyacrylamidegels (BioRad, Watford, UK) and transferred onto an Immobilonnitrocellulose membrane (Millipore). Non-specific binding was blockedwith 1% casein (Vector Laboratories) before incubation with rabbitmonoclonal antibodies to human N-cadherin (D4R1H) at a dilution of1:1000, E-cadherin (24E10) at a dilution of 1:500 or gamma-catenin(2303) at a dilution of 1:500 (Cell signalling) or mouse monoclonalGAPDH (ab8245) at a dilution of 1:1000 (AbCam, Cambridge UK) for 16 h at4° C. Secondary antibodies were anti-rabbit or anti-mouse horse radishperoxidase (HRP; 1:15,000) and HRP was detected with the Supersignalchemiluminescence detection kit (Pierce). Band quantification wascarried out using Quantity Once software (BioRad) and normalised toGAPDH.

Gene Analysis

Total RNA was extracted using an RNeasy kit (Qiagen) and reversetranscribed into cDNA using Superscript III (Invitrogen AB). RelativemRNA expression of IL-1B (Hs02786624), IL-1R1 (Hs00174097), CASP(Caspase 1) (Hs00354836), IL1RN (Hs00893626), JUP (junctionplakoglobin/gamma-catenin) (Hs00984034), N-cadherin (Hs01566408) andE-cadherin (Hs1013933) were compared with the housekeeping geneglyceraldehyde-3-phosphate dehydrogenase (GAPDH; Hs02786624) andassessed using an ABI 7900 PCR System (Perkin Elmer, Foster City,Calif.) and Taqman universal master mix (Thermofisher, UK). Fold changein gene expression between treatment groups was analysed by inserting CTvalues into Data Assist V3.01 software (Applied Biosystems) and changesin gene expression were only analysed for genes with a CT value of ≤25.

Assessment of IL-1β and IL-1R1 in Tumors from Breast Cancer Patients

IL-1β and IL-1R1 expression was assessed on tissue microarrays (TMA)containing primary breast tumor cores taken from 1,300 patients includedin the clinical trial, AZURE (Coleman et al. 2011). Samples were takenpre-treatment from patients with stage II and III breast cancer withoutevidence of metastasis. Patients were subsequently randomized tostandard adjuvant therapy with or without the addition of zoledronicacid for 10 years (Coleman et al 2011). The TMAs were stained for IL-1β(ab2105, 1:200 dilution, Abcam) and IL-1R1 (ab59995, 1:25 dilution,Abcam) and scored blindly under the guidance of a histopathologist forIL-1β/IL-1R1 in the tumor cells or in the associated stroma. Tumor orstromal IL-1β or IL-1R1 was then linked to disease recurrence (any site)or disease recurrence specifically in bone (+/− other sites).

The IL-1β Pathway is Upregulated During the Process of Human BreastCancer Metastasis to Human Bone.

A mouse model of spontaneous human breast cancer metastasis to humanbone implants was utilised to investigate how the IL-1β pathway changesthrough the different stages of metastasis. Using this model, theexpression levels of genes associated with the IL-1β pathway increasedin a stepwise manner at each stage of the metastatic process in bothtriple negative (MDA-MB-231) and estrogen receptor positive (ER+ve)(T47D) breast cancer cells: Genes associated with the IL-1β signallingpathway (IL-1B, IL-1R1, CASP (Caspase 1) and IL-1Ra) were expressed atvery low levels in both MDA-MB-231 and T47D cells grown in vitro andexpression of these genes were not altered in primary mammary tumorsfrom the same cells that did not metastasize in vivo (FIG. 1a ).

IL-1B, IL-1R1 and CASP were all significantly increased in mammarytumors that subsequently metastasized to human bone compared with thosethat did not metastasize (p<0.01 for both cell lines), leading toactivation of IL-1β signalling as shown by ELISA for the active 17 kDIL-1β (FIG. 1 b; FIG. 2). IL-1B gene expression increased in circulatingtumor cells compared with metastatic mammary tumors (p<0.01 for bothcell lines) and IL-1B (p<0.001), IL-1R1 (p<0.01), CASP (p<0.001) andIL-1Ra (p<0.01) were further increased in tumor cells isolated frommetastases in human bone compared with their corresponding mammarytumors, leading to further activation of IL-1β protein (FIG. 1; FIG. 2).These data suggest that IL-1β signalling may promote both initiation ofmetastasis from the primary site as well as development of breast cancermetastases in bone.

Tumor Derived IL-1β Promotes EMT and Breast Cancer Metastasis.

Expression levels of genes associated with tumor cell adhesion andepithelial to mesenchymal transition (EMT) were significantly altered inprimary tumors that metastasized to bone compared with tumors that didnot metastasize (FIG. 1c ). IL-1β-overexpressing cells were generated(MDA-MB-231-IL-1B+, T47D-IL-1B+ and MCF7-IL-1B+) to investigate whethertumor-derived IL-1β is responsible for inducing EMT and metastasis tobone. All IL-1β+ cell lines demonstrated increased EMT exhibitingmorphological changes from an epithelial to mesenchymal phenotype (FIG.3a ) as well as reduced expression of E-cadherin, and JUP (junctionplakoglobin/gamma-catenin) and increased expression of N-Cadherin geneand protein (FIG. 3b ). Wound closure (p<0.0001 in MDA-MB-231-IL-1β+(FIG. 3d ); p<0.001 MCF7-IL-1β+ and T47D-IL-1β+) and migration andinvasion through matrigel towards osteoblasts were increased in tumorcells with increased IL-1β signalling compared with their respectivecontrols (MDA-MB-231-IL-1β+ (FIG. 3c ) p<0.0001; MCF7-IL-1β+ andT47D-IL-1β+ p<0.001). Increased IL-1β production was seen in ER-positiveand ER-negative breast cancer cells that spontaneously metastasized tohuman bone implants in vivo compared with non-metastatic breast cancercells (FIG. 1). The same link between IL-1β and metastasis was made inprimary tumor samples from patients with stage II and III breast cancerenrolled in the AZURE study (Coleman et al., 2011) that experiencedcancer relapsed over a 10 year time period. IL-1β expression in primarytumors from the AZURE patients correlated with both relapse in bone andrelapse at any site indicating that presence of this cytokine is likelyto play a role in metastasis in general. In agreement with this, geneticmanipulation of breast cancer cells to artificially overexpress IL-1βincreased the migration and invasion capacities of breast cancer cellsin vitro (FIG. 3).

Inhibition of IL-1β Signaling Reduces Spontaneous Metastasis to HumanBone.

As tumor derived IL-1β appeared to be promoting onset of metastasisthrough induction of EMT the effects of inhibiting IL-1β signaling withIL-1Ra (Anakinra) or a human anti-IL-1β-binding antibody (canakinumab)on spontaneous metastasis to human bone implants were investigated: BothIL-1Ra and canakinumab reduced metastasis to human bone: metastasis wasdetected in human bone implants in 7 out of 10 control mice, but only in4 out of 10 mice treated with IL-1Ra and 1 out of 10 mice treated withcanakinumab. Bone metastases from IL-1Ra and canakinumab treatmentgroups were also smaller than those detected in the control group (FIG.4a ). Numbers of cells detected in the circulation of mice treated withcanakinumab or IL-1Ra were significantly lower than those detected inthe placebo treated group: 3 and 3 tumor cells/ml were counted in wholeblood from mice treated with canakinumab and anakinra, respectively,compared 108 tumor cells/ml counted in blood from placebo treated mice(FIG. 4b ), suggesting that inhibition of IL-1 signalling prevents tumorcells from being shed from the primary site into the circulation.Therefore, inhibition of IL-1β signaling with the anti-IL-1β antibodycanakinumab or inhibition of IL-1R1 reduced the number of breast cancercells shed into the circulation and reduced metastases in human boneimplants (FIG. 4).

Tumor Derived IL-1B Promotes Bone Homing and Colonisation of BreastCancer Cells.

Injection of breast cancer cells into the tail vein of mice usuallyresults in lung metastasis due to the tumor cells becoming trapped inthe lung capillaries. It was previously shown that breast cancer cellsthat preferentially home to the bone microenvironment followingintra-venous injection express high levels of IL-1β, suggesting thatthis cytokine may be involved in tissue specific homing of breast cancercells to bone. In the current study, intravenous injection ofMDA-MB-231-IL-1β+ cells into BALB/c nude mice resulted in significantlyincreased number of animals developing bone metastasis (75%) comparedwith control cells (12%) (p<0.001) cells (FIG. 5a ). MDA-MB-231-IL-1β+tumors caused development of significantly larger osteolytic lesions inmouse bone compared with control cells (p=0.03; FIG. 5b ) and there wasa trend towards fewer lung metastases in mice injected withMDA-MB-231-IL-1β+ cells compared with control cells (p=0.16; FIG. 5c ).These data suggest that endogenous IL-1β can promote tumor cell homingto the bone environment and development of metastases at this site.

Tumor Cell-Bone Cell Interactions Further Induce IL-1B and PromoteDevelopment of Overt Metastases.

Gene analysis data from a mouse model of human breast cancer metastasisto human bone implants suggested that the IL-1β pathway was furtherincreased when breast cancer cells are growing in the bone environmentcompared with metastatic cells in the primary site or in the circulation(FIG. 1a ). It was therefore investigated how IL-1β production changeswhen tumor cells come into contact with bone cells and how IL-1β altersthe bone microenvironment to affect tumor growth (FIG. 6). Culture ofhuman breast cancer cells into pieces of whole human bone for 48 hresulted in increased secretion of IL-1β into the medium (p<0.0001 forMDA-MB-231 and T47D cells; FIG. 6a ). Co-culture with human HS5 bonemarrow cells revealed the increased IL-1β concentrations originated fromboth the cancer cells (p<0.001) and bone marrow cells (p<0.001), withIL-1β from tumor cells increasing ˜1000 fold and IL-1B from HS5 cellsincreasing ˜100 fold following co-culture (FIG. 6b ).

Exogenous IL-1β did not increase tumor cell proliferation, even in cellsoverexpressing IL-1R1. Instead, IL-1β stimulated proliferation of bonemarrow cells, osteoblasts and blood vessels that in turn inducedproliferation of tumor cells (FIG. 5). It is therefore likely thatarrival of tumor cells expressing high concentrations of IL-1β stimulateexpansion of the metastatic niche components and contact between IL-1βexpressing tumor cells and osteoblasts/blood vessels drive tumorcolonization of bone. The effects of exogenous IL-1β as well as IL-1βfrom tumor cells on proliferation of tumor cells, osteoblasts, bonemarrow cells and CD34⁺ blood vessels were investigated: Co-culture ofHS5 bone marrow or OB1 primary osteoblast cells with breast cancer cellscaused increased proliferation of all cell types (P<0.001 for HS5,MDA-MB-231 or T47D, FIG. 6c ) (P<0.001 for OB1, MDA-MB-231 or T47D, FIG.6d ). Direct contact between tumor cells, primary human bone samples,bone marrow cells or osteoblasts promoted release of IL-1β from bothtumor and bone cells (FIG. 6). Furthermore, administration of IL-1βincreased proliferation of HS5 or OB1 cells but not breast cancer cells(FIGS. 7a and b ), suggesting that tumor cell-bone cell interactionspromote production of IL-1β that can drive expansion of the niche andstimulate the formation of overt metastases.

IL-1β signalling was also found to have profound effects on the bonemicrovasculature: Preventing IL-1β signaling in bone by knocking outIL-1R1, pharmacological blockade of IL-1R with IL-1Ra or reducingcirculating concentrations of IL-1β by administering the anti-IL-1βbinding antibody canakinumab reduced the average length of CD34⁺ bloodvessels in trabecular bone, where tumor colonisation takes place (p<0.01for IL-1Ra and canakinumab treated mice) (FIG. 7c ). These findings wereconfirmed by endomeucin staining which showed decreased numbers of bloodvessels as well as blood vessel length in bone when IL-1β signaling wasdisrupted. ELISA analysis for endothelin 1 and VEGF showed reducedconcentrations of both of these endothelial cell markers in the bonemarrow for IL-1R1^(−/−) mice (p<0.001 endothelin 1; p<0.001 VEGF) andmice treated with IL-1R antagonist (p<0.01 endothelin 1; p<0.01 VEGF) orcanakinumab (p<0.01 endothelin 1; p<0.001 VEGF) compared with control(FIG. 8). These data suggest that tumor cell-bone cell associatedincreases in IL-1β and high levels of IL-1β in tumor cells may alsopromote angiogenesis, further stimulating metastases.

Tumor Derived IL-1β Predicts Future Breast Cancer Relapse in Bone andOther Organs in Patient Material

To establish the relevance of the findings in a clinical setting thecorrelation between IL-1β and its receptor IL-1R1 in patient samples wasinvestigated. ˜1300 primary tumor samples from patients with stageII/III breast cancer with no evidence of metastasis (from the AZUREstudy (Coleman et al., 2011)) were stained for IL-1R1 or the active (17kD) form of IL-1β, and biopsies were scored separately for expression ofthese molecules in the tumor cells and the tumor associated stroma.Patients were followed up for 10 years following biopsy and correlationbetween IL-1β/IL-1R1 expression and distant recurrence or relapse inbone assessed using a multivariate Cox model. IL-1β in tumor cellsstrongly correlated with distant recurrence at any site (p=0.0016),recurrence only in bone (p=0.017) or recurrence in bone at any time(p=0.0387) (FIG. 9). Patients who had IL-1β in their tumor cells andIL-1R1 in the tumor associated stroma were more likely to experiencefuture relapse at a distant site (p=0.042) compared to patients who didnot have IL-1β in their tumor cells, indicating that tumor derived IL-1βmay not only promote metastasis directly but may also interact withIL-1R1 in the stroma to promote this process. Therefore, IL-1β is anovel biomarker that can be used to predict risk of breast cancerrelapse.

Example 2

Simulation of Canakinumab PK Profile and hsCRP Profile for Lung CancerPatients.

A model was generated to characterize the relationship betweencanakinumab pharmacokinetics (PK) and hsCRP based on data from theCANTOS study.

The following methods were used in this study: Model building wasperformed using the first-order conditional estimation with interactionmethod. The model described the logarithm of the time resolved hsCRP as:

y(t _(ij))=y _(0, i) +y _(eff)(t _(ij))

where y_(0, i) is a steady state value and y_(eff)(t_(ij)) describes theeffect of the treatment and depends on the systemic exposure. Thetreatment effect was described by an Emax-type model,

${y_{eff}\left( t_{ij} \right)} = {E_{\max,i}\frac{c\left( t_{ij} \right)}{{c\left( t_{ij} \right)} + {{IC}\; 50_{i}}}}$

where E_(max, i) is the maximal possible response at high exposure, andIC50_(i) is the concentration at which half maximal response isobtained.

The individual parameters, E_(max, i) and y_(0, i) and the logarithm ofIC50_(i) were estimated as a sum of a typical value, covariate effectscovpar*cov_(i) and normally distributed between subject variability. Inthe term for the covariate effect covpar refers to the covariate effectparameter being estimated and cov_(i) is the value of the covariate ofsubject i. Covariates to be included were selected based on inspectionof the eta plots versus covariates. The residual error was described asa combination of proportional and additive term.

The logarithm of baseline hsCRP was included as covariate on all threeparameters (E_(max, i), y_(0, i) and IC50_(i)). No other covariate wasincluded into the model. All parameters were estimated with goodprecision. The effect of the logarithm of the baseline hsCRP on thesteady state value was less than 1 (equal to 0.67). This indicates thatthe baseline hsCRP is an imperfect measure for the steady state value,and that the steady state value exposes regression to the mean relativeto the baseline value. The effects of the logarithm of the baselinehsCRP on IC50 and Emax were both negative. Thus patients with high hsCRPat baseline are expected to have low IC50 and large maximal reductions.In general, model diagnostics confirmed that the model describes theavailable hsCRP data well.

The model was then used to simulate expected hsCRP response for aselection of different dosing regimens in a lung cancer patientpopulation. Bootstrapping was applied to construct populations withintended inclusion/exclusion criteria that represent potential lungcancer patient populations. Three different lung cancer patientpopulations described by baseline hsCRP distribution alone wereinvestigated: all CANTOS patients (scenario 1), confirmed lung cancerpatients (scenario 2), and advanced lung cancer patients (scenario 3).

The population parameters and inter-patient variability of the modelwere assumed to be the same for all three scenarios. The PK/PDrelationship on hsCRP observed in the overall CANTOS population wasassumed to be representative for lung cancer patients.

The estimator of interest was the probability of hsCRP at end of month 3being below a cut point, which could be either 2 mg/L or 1.8 mg/L. 1.8mg/L was the median of hsCRP level at end of month 3 in the CANTOSstudy. Baseline hsCRP >2 mg/L was one of the inclusion criteria, so itis worthy to explore if hsCRP level at end of month 3 went below 2 mg/L.

A one-compartment model with first order absorption and elimination wasestablished for CANTOS PK data. The model was expressed as ordinarydifferential equation and R×ODE was used to simulate canakinumabconcentration time course given individual PK parameters. Thesubcutaneous canakinumab dose regimens of interest were 300 mg Q12W, 200mg Q3W, and 300 mg Q4W. Exposure metrics including Cmin, Cmax, AUCs overdifferent selected time periods, and average concentration Cave atsteady state were derived from simulated concentration time profiles.

The simulation in Scenario 1 was based on the below information:

Individual canakinumab exposure simulated using R×ODE

PD parameters which are components of y_(0, i), E_(max, i), andIC50_(i): typical values (THETA(3), THETA(5), THETA(6)), covpars(THETA(4), THETA(7), THETA(8)), and between subject variability (ETA(1),ETA(2), ETA(3))

Baseline hsCRP from all 10,059 CANTOS study patients (baseline hsCRP:mean 6.18 mg/L, standard error of the mean (SEM)=0.10 mg/L)

The prediction interval of the estimator of interest was produced byfirst randomly sampling 1000 THETA(3)-(8)s from a normal distributionwith fixed mean and standard deviation estimated from the populationPK/PD model; and then for each set of THETA(3)-(8), bootstrapping 2000PK exposure, PD parameters ETA(1)-(3), and baseline hsCRP from allCANTOS patients. The 2.5%, 50%, and 97.5% percentile of 1000 estimateswere reported as point estimator as well as 95% prediction interval.

The simulation in Scenario 2 was based on the below information:

Individual canakinumab PK exposure simulated using R×ODE

PD parameters THETA(3)-(8) and ETA(1)-(3)

Baseline hsCRP from 116 CANTOS patients with confirmed lung cancer(baseline hsCRP: mean=9.75 mg/L, SEM=1.14 mg/L)

The prediction interval of the estimator of interest was produced byfirst randomly sampling 1000 THETA(3)-(8)s from a normal distributionwith fixed mean and standard deviation estimated from the populationPKPD model; and then for each set of THETA(3)-(8), bootstrapping 2000 PKexposure, PD parameters ETA(1)-(3) from all CANTOS patients, andbootstrapping 2000 baseline hsCRP from the 116 CANTOS patients withconfirmed lung cancer. The 2.5%, 50%, and 97.5% percentile of 1000estimates were reported as point estimator as well as 95% predictioninterval.

In Scenario 3, the point estimator and 95% prediction interval wereobtained in a similar manner as for scenario 2. The only difference wasbootstrapping 2000 baseline hsCRP values from advanced lung cancerpopulation. There is no individual baseline hsCRP data published in anadvanced lung cancer population. An available population level estimatein advanced lung cancer is a mean of baseline hsCRP of 23.94 mg/L withSEM 1.93 mg/L [Vaguliene 2011].

Using this estimate, the advanced lung cancer population was derivedfrom the 116 CANTOS patients with confirmed lung cancer using anadditive constant to adjust the mean value to 23.94 mg/L.

In line with the model, the simulated canakinumab PK was linear. Themedian and 95% prediction interval of concentration time profiles areplotted in natural logarithm scale over 6 months is shown in FIG. 10 a.

The median and 95% prediction intervals of 1000 estimates of proportionof subjects with month 3 hsCRP response under the cut point of 1.8 mg/Land 2 mg/L mhsCRP are reported in FIGS. 10b and c. Judging from thesimulation data, 200 mg Q3W and 300 mg Q4W perform similarly and betterthan 300 mg Q12W (top dosing regimen in CANTOS) in terms of decreasinghsCRP at month 3. Going from scenario 1 to scenario 3 towards moresevere lung cancer patients, higher baseline hsCRP levels are assumed,and result in smaller probabilities of month 3 hsCRP being below the cutpoint. FIG. 10d shows how the median hsCRP concentration changes overtime for three different doses and FIG. 10e shows the percent reductionfrom baseline hsCRP after a single dose.

Example 3 PDR001 Plus Canakinumab Treatment Increases EffectorNeutrophils in Colorectal Tumors.

RNA sequencing was used to gain insights on the mechanism of action ofcanakinumab (ACZ885) in cancer. The CPDR001X2102 and CPDR001X2103clinical trials evaluate the safety, tolerability and pharmacodynamicsof spartalizumab (PDR001) in combination with additional therapies. Foreach patient, a tumor biopsy was obtained prior to treatment, as well ascycle 3 of treatment. In brief, samples were processed by RNAextraction, ribosomal RNA depletion, library construction andsequencing. Sequence reads were aligned by STAR to the hg19 referencegenome and Refseq reference transcriptome, gene-level counts werecompiled by HTSeq, and sample-level normalization using the trimmed meanof M-values was performed by edgeR.

FIG. 11 shows 21 genes that were increased, on average, in colorectaltumors treated with PDR001+canakinumab (ACZ885), but not in colorectaltumors treated with PDR001+everolimus (RAD001). Treatment withPDR001+canakinumab increased the RNA levels of IL1B, as well as itsreceptor, IL1R2. This observation suggests an on-target compensatoryfeedback by tumors to increase IL1B RNA levels in response to IL-1βprotein blockade.

Notably, several neutrophil-specific genes were increased onPDR001+canakinumab, including FCGR3B, CXCR2, FFAR2, OSM, and G0S2(indicated by boxes in FIG. 11). The FCGR3B gene is aneutrophil-specific isoform of the CD16 protein. The protein encoded byFCGR3B plays a pivotal role in the secretion of reactive oxygen speciesin response to immune complexes, consistent with a function of effectorneutrophils (Fossati G 2002 Arthritis Rheum 46: 1351). Chemokines thatbind to CXCR2 mobilize neutrophils out of the bone marrow and intoperipheral sites. In addition, increased CCL3 RNA was observed ontreatment with PDR001+canakinumab. CCL3 is a chemoattractant forneutrophils (Reichel C A 2012 Blood 120: 880).

In summary, this contribution of components analysis using RNA-seq datademonstrates that PDR001+canakinumab treatment increases effectorneutrophils in colorectal tumors, and that this increase was notobserved with PDR001+everolimus treatment.

Example 4

Efficacy of Canakinumab (ACZ885) in Combination with Spartalizumab(PDR001) in the Treatment of Cancer.

Patient 5002-004 is a 56 year old man with initially Stage IIC,microsatellite-stable, moderately differentiated adenocarcinoma of theascending colon (MSS-CRC), diagnosed in June, 2012 and treated withprior regimens.

Prior treatment regimens included:

1. Folinic acid/5-fluorouracil/oxaliplatin in the adjuvant setting

2. Chemoradiation with capecitabine (metastatic setting)

3. 5-fluorouracil/bevacizumab/folinic acid/irinotecan

4. trifluridine and tipiracil

5. Irinotecan

6. Oxaliplatin/5-fluorouracil

7. 5-fluorouracil/bevacizumab/leucovorin

8. 5-fluorouracil

At study entry the patient had extensive metastatic disease includingmultiple hepatic and bilateral lung metastases, and disease inparaesophageal lymph nodes, retroperitoneum and peritoneum.

The patient was treated with PDR001 400 mg every four weeks (Q4W) plus100 mg every eight weeks (Q8W) ACZ885. The patient had stable diseasefor 6 months of therapy, then with substantial disease reduction andconfirmed RECIST partial response to treatment at 10 months. The patienthas subsequently developed progressive disease and the dose wasincreased to 300 mg and then to 600 mg.

Example 5 Calculations for Selecting the Dose for Gevokizumab for CancerPatients.

Dose selection for gevokizumab in the treatment of cancer having atleast partial inflammatory basis is based on the clinical effectivedosings reveals by the CANTOS trial in combination with the available PKdata of gevokizumab, taking into the consideration that Gevokizumab(IC50 of ˜2-5 pM) shows a ˜10 times higher in vitro potency compared tocanakinumab (IC50 of ˜42±3.4 pM). The gevokizumab top dose of 0.3 mg/kg(˜20 mg) Q4W showed reduction of hsCRP could reduce hsCRP up to 45% intype 2 diabetes patients (see FIG. 12a ).

Next, a pharmacometric model was used to explore the hsCRPexposure-response relationship, and to extrapolate the clinical data tohigher ranges. As clinical data show a linear correlation between thehsCRP concentration and the concentration of gevokizumab (both inlog-space), a linear model was used. The results are shown in FIG. 12 b.Based on that simulation, a gevokizumab concentration between 10000ng/mL and 25000 ng/mL is optimal because hsCRP is greatly reduced inthis range, and there is only a diminishing return with gevokizumabconcentrations above 15000 ng/mL. However gevokizumab concentrationsbetween 4000 ng/mL and 10000 ng/mL is expected to be efficacious ashsCRP has already been significantly reduced in that range.

Clinical data showed that gevokizumab pharmacokinetics follow a lineartwo-compartment model with first order absorption after a subcutaneousadministration. Bioavailability of gevokizumab is about 56% whenadministered subcutaneously. Simulation of multiple-dose gevokizumab(SC) was carried out for 100 mg every four weeks (see FIG. 12c ) and 200mg every four weeks (see FIG. 12d ). The simulations showed that thetrough concentration of 100 mg gevokizumab given every four weeks isabout 10700 ng/mL. The half-life of gevokizumab is about 35 days. Thetrough concentration of 200 mg gevokizumab given every four weeks isabout 21500 ng/mL.

Example 6 Preclinical Data on the Effects of Anti-IL-1beta Treatment

Canakinumab, an anti-IL-1β human IgG1 antibody, cannot directly beevaluated in mouse models of cancer due to the fact that it does notcross-react with mouse IL-1β. A mouse surrogate anti-IL-1β antibody hasbeen developed and is being used to evaluate the effects of blockingIL-1β in mouse models of cancer. This isotype of the surrogate antibodyis IgG2a, which is closely related to human IgG1.

In the MC38 mouse model of colon cancer, modulation of tumorinfiltrating lymphocytes (TILs) can be seen after one dose of the antiIL-1β antibody (FIG. 13a-c ). MC38 tumors were subcutaneously implantedin the flank of C57BL/6 mice and when the tumors were between 100-150mm³, the mice were treated with one dose of either an isotype antibodyor the anti IL-1β antibody. Tumors were then harvested five days afterthe dose and processed to obtain a single cell suspension of immunecells. The cells were then ex vivo stained and analyzed via flowcytometry. Following a single dose of an IL-1β blocking antibody, thereis an increase in in CD4+ T cells infiltrating the tumor and also aslight increase in CD8+ T cells (FIG. 13a ).

The CD8+ T cell increase is slight but may allude to a more activeimmune response in the tumor microenvironment, which could potentiallybe enhanced with combination therapies. The CD4+ T cells were furthersubdivided into FoxP3+ regulatory T cells (Tregs), and this subsetdecreases following blockade of IL-1β (FIG. 13b ). Among the myeloidcell populations, blockade of IL-1β results in a decrease in neutrophilsand the M2 subset of macrophages, TAM2 (FIG. 13c ). Both neutrophils andM2 macrophages can be suppressive to other immune cells, such asactivated T cells (Pillay et al, 2013; Hao et al, 2013; Oishi et al2016). Taken together, the decrease in Tregs, neutrophils, and M2macrophages, in the MC38 tumor microenvironment following IL-1β blockadeargues that the tumor microenvironment is becoming less immunesuppressive.

In the LL2 mouse model of lung cancer, a similar trend towards a lesssuppressive immune microenvironment can be seen after one dose of ananti-IL-1β antibody (FIG. 13d-f ). LL2 tumors were subcutaneouslyimplanted in the flank of C57BL/6 mice and when the tumors were between100-150 mm3, the mice were treated with one dose of either an isotypeantibody or the anti-IL-1β antibody. Tumors were then harvested fivedays after the dose and processed to obtain a single cell suspension ofimmune cells. The cells were then ex vivo stained and analyzed via flowcytometry. There is a decrease in the Treg populations as evaluated bythe expression of FoxP3 and Helios (FIG. 13d ). FoxP3 and Helios areboth used as markers of regulatory T cells, while they may definedifferent subsets of Tregs (Thornton et al, 2016).

Similar to the MC38 model, there is a decrease in both neutrophils andM2 macrophages (TAM2) following IL-1β blockade (FIG. 13e ). In additionto this, in this model the change in the myeloid derived suppressor cell(MDSC) populations were evaluated following antibody treatment. Thegranulocytic or polymorphonuclear (PMN) MDSC were found in reducednumbers following anti-IL-1β treatment (FIG. 13f ). MDSC are a mixedpopulation of cells of myeloid origin that can actively suppress T cellresponses through several mechanisms, including arginase production,reactive oxygen species (ROS) and nitric oxide (NO) release (Kumar etal, 2016; Umansky et al, 2016). Again, the decrease in Tregs,neutrophils, M2 macrophages, and PMN MDSC in the LL2 model followingIL-1β blockade argues that the tumor microenvironment is becoming lessimmune suppressive.

TILs in the 4T1 triple negative breast cancer model also show a trendtowards a less suppressive immune microenvironment after one dose of themouse surrogate anti-IL-1β antibody (FIG. 13g-j ). 4T1 tumors weresubcutaneously implanted in the flank of Balb/c mice, and the mice weretreated with either an isotype antibody or the anti-IL-1β antibody whenthe tumors were between 100-150 mm3. Tumors were then harvested fivedays after the dose and processed to obtain a single cell suspension ofimmune cells. The cells were then ex vivo stained and analyzed via flowcytometry. There is a decrease in CD4+ T cells after a single dose of ananti-IL-1β antibody (FIG. 13g ) and within the CD4+ T cell population,there is a decrease in the FoxP3+ Tregs (FIG. 13h ). Further, there is adecrease in both the TAM2 and neutrophil populations following treatmentof the tumor-bearing mice (FIG. 13i ). All of these data together againargue that IL-1β blockade in the 4T1 breast cancer mouse model leads toa less suppressive immune microenvironment. In addition to this, in thismodel the MDSC populations was also evaluated following antibodytreatment. Both the granulocytic (PMN) MDSC and monocytic MDSC werefound in reduced numbers following anti-IL-1β treatment (FIG. 13j ).These findings in combination with the changes in Tregs, M2 macrophages,and the neutrophil populations describe a decrease in the immunesuppressive tumor microenvironment in the 4T1 tumor model.

While these data are from colon, lung, and breast cancer models, thedata can be extrapolated to other types of cancer. Even though thesemodels do not fully correlate to human cancers of the same type, theMC38 model in particular is a good surrogate model for hypermutated/MSI(microsatellite instable) colorectal cancer (CRC). Based on thetranscriptomic characterization of the MC38 cell line, four of thedriver mutations in this line correspond to known hotspots in human CRC,although these are at different positions (Efremova et al, 2018). Whilethis does not make the MC38 mouse model identical to human CRC, it doesmean that MC38 may be a relevant model for human MSI CRC. Generally,mouse models do not always correlate to the same type of cancer inhumans due to genetic differences in the origins of the cancer in miceversus humans. However, when examining the infiltrating immune cells,the type of cancer is not always important, as the immune cells are morerelevant. In this case, as three different mouse models show a similardecrease in the suppressive microenvironment of the tumor, blockingIL-1β seems to lead to a less suppressive tumor microenvironment. Theextent of the change in immune suppression with multiple cell types(Tregs, TAMs, neutrophils) showing a decrease compared to the isotypecontrol in multiple tumor syngeneic mouse tumor models is a novelfinding for IL-1β blockade in mouse models of cancer. While suppressorcell decreases have been seen before, multiple cell types in each modelis a novel finding. In addition, changes to MDSC populations in the 4T1and Lewis lung carcinoma (LL2) models have been seen downstream ofIL-1β, but the finding in the LL2 model that blockade of IL-1β can leadto the reduction of MDSCs is novel to this study and the mouse surrogateof canakinumab (Elkabets et al, 2010).

Even though these models do not fully correlate to human cancers of thesame type, the MC38 model in particular is a good surrogate model forhypermutated/MSI (microsatellite instable) colorectal cancer (CRC).Based on the transcriptomic characterization of the MC38 cell line, fourof the driver mutations in this line correspond to known hotspots inhuman CRC, although these are at different positions (Efremova et al,2018). While this does not make the MC38 mouse model identical to humanCRC, it does mean that MC38 may be a relevant model for human MSI CRC(Efremova M, et al. Nature Communications 2018; 9: 32).

Example 7

Preclinical Data on the Efficacy of Canakinumab in Combination with anAnti-PD-1 (Pembrolizumab) in the Treatment of Cancer.

A pilot study was designed to assess the impact of canakinumab as amonotherapy or in combination with anti-PD-1 (pembrolizumab) on tumorgrowth and the tumor microenvironment. A xenograft model of human NSCLCwas created by subcutaneous injection of a human lung cancer cell lineH358 (KRAS mutant) into BLT mouse xenograft model.

As shown in FIG. 14, the H358 (KRAS mutant) model is a very fast growingand aggressive model. In this model, combination treatment ofcanakinumab and pembrolizumab (shown in purple) led to a greaterreduction than canakinumab single agent arm (shown in red) andpembrolizumab single agent treatment (shown in green), with a 50%decrease observed in the mean tumor volume when compared to the vehiclegroup.

Example 8

Preclinical Data on the Efficacy of Canakinumab in Combination withDocetaxel in the Treatment of Cancer.

In a study of anti-IL-1β in combination with docetaxel in an aggressivelung model (LL2), modest efficacy with anti-IL-1β was observed, as wellas docetaxel alone. The efficacy was enhanced in the combinationcompared to either group alone or control (FIG. 15A). Decreases inimmunosuppressive cells were observed with anti-IL-1β alone or incombination at the PD time point 5 days after the first dose,specifically in regulatory T cells and suppressive mouse myeloid cellsincluding neutrophils, monocytes and MDSCs in tumors after IL-1βinhibition (FIG. 15B-E). These data support that the proposed mechanismof action in IL-1β inhibition can be demonstrated in vivo and also someefficacy of anti-IL-1β monotherapy was observed.

Example 9

Treatment of 4T1 Tumors with 01BSUR and Docetaxel Leads to Alterationsin the Tumor Microenvironment.

Female Balb/c mice with 4T1 tumors implanted subcutaneously (s.c.) onthe right flank were treated 8 and 15 days post-tumor implant initiatingwhen the tumors reached about 100 mm³ with the isotype antibody,docetaxel, 01BSUR, or a combination of docetaxel and 01BSUR. 01BSUR isthe mouse surrogate antibody, since canakinumab does not cross-react tomurine IL-1beta. 01BSUR belongs to the mouse IgG2a subclass, whichcorresponds to human IgG1 subclass, which canakinumab belongs to. 5 daysafter the first dose, tumors were harvested and analyzed for changes tothe infiltrating immune cell populations. This was done again at the endpoint of the study, 4 days after the second dose.

Tumor Burden

A slight slowing in tumor growth was seen in the 01BSUR anti-IL-1β alonetreatment group compared to the vehicle/isotype control. This delay wasenhanced in the single agent docetaxel group. The combination groupshowed a similar slowing in growth as the docetaxel alone group (FIG.16).

TIL Analysis of 4T1 Tumors After a Single Dose of Docetaxel and01BSUR—Myeloid Panel

Following a single treatment with docetaxel alone or in combination with01BSUR, there was a decrease in neutrophils in the 4T1 tumors. Thecombination group, showed a greater decrease in neutrophil cell numberthan the docetaxel single agent group. Single agent 01BSUR led to aslight increase in neutrophils in 4T1 tumors, although this was not asignificant change compared to the control group. Each of the treatmentsled to a decrease in monocytes compared to the vehicle/isotype group.The single agent 01BSUR treatment led to a greater decrease in monocytesthan the docetaxel alone group. Further, the combination showed an evengreater decrease in monocytes compared to the control group (P=0.0481)(FIG. 17). Similar trends to the granulocytes and monocytes were seenamong the granulocytic and monocytic Myeloid derived suppressor cells(MDSC). Docetaxel alone and in combination with 01BSUR led to a decreasein granulocytic MDSC. All treatments led to a decrease in monocyticMDSC, with the combination leading to a greater decrease than either ofthe single agents (FIG. 18).

TIL Analysis of 4T1 Tumors After a Second Dose of Docetaxel and 01BSUR

Four days after a second dose of docetaxel and 01BSUR 4T1 tumors wereanalyzed for immune cell infiltrates. The percent of both CD4⁺ and CD8⁺T cells expressing TIM-3 were determined. Docetaxel alone led to nochange in the TIM-3 expressing cells compared to the control group,while there was a decrease in the TIM-3 expressing cells followingtreatment with 01BSUR alone or in combination with docetaxel. Thecombination group, appears to show a slightly larger decrease in TIM-3expressing cells than the single agent 01BSUR group (P=0.0063) for CD4⁺T cells compared to control (FIG. 19). Similar trends were seen in theTreg subset of cells with the combination group showing the largestlevel of decrease of the TIM-3 expressing cells (P=0.0064) compared tothe control (FIG. 20).

Conclusion and Discussion

Blocking IL-1β has been shown to be a potent method of changing theinflammatory microenvironment in autoimmune disease. ACZ885(canakinumab) has been highly effective at treating some inflammatoryautoimmune diseases, such as CAPS (Cryopyrin Associated PeriodicSyndrome). As many tumors have an inflammatory microenvironment,blocking IL-1β is being studied to determine the impact that this willhave on the tumor microenvironment alone and in combination with agentsthat will work to block the PD-1/PD-L1 axis or standard of carechemotherapeutic agents such as docetaxel. It has been shown throughpreclinical experiments and the CANTOS trial that the blockade of IL-1βcan have an impact on tumor growth and development. However, the CANTOStrial, an atherosclerosis trial, evaluated this in a prophylacticsetting with patients with no known or detectable cancer at the time ofenrollment. Patients with established tumors or metastases may havedifferent levels of response to IL-1β blockade.

These preliminary results studying combinations of 01BSUR, a murinesurrogate of ACZ885, and docetaxel show that in the LL2 and 4T1 tumorsmodels, this combination can have an impact on tumor growth.

The studies described here examine the TILs following a single treatmentonly (1D2 and 01BSUR combinations) or following two doses of eachtreatment (01BSUR and docetaxel).

The overall trends alludes to a change in the suppressive nature of theTME in LL2 and 4T1 tumors.

While there is not a consistent change in the overall CD4⁺ and CD8⁺ Tcells in the TME of these tumors, there is a trend towards in decreasein the Tregs in these tumors. Additionally, the Tregs typically alsoshow a decrease in the percentage of cells expressing TIM-3. Tregs thatexpress TIM-3 may be more effective suppressors of T cells thannon-TIM-3 expressing Tregs [Sakuishi, 2013]. In several of the studies,there is an overall decrease of TIM-3 on all T cells.

While the impact of this on these cells is not yet known, TIM-3 is acheckpoint and these cells may be more activated than the TIM-3expressing T cells. However, further work is needed to understand thesechanges as some of the T cell changes observed could allude to a therapythat is less effective than the control.

While T cells make up a portion of the immune cell infiltrate in thesetumors, a large portion of the infiltrating cells are myeloid cells.These cells were also analyzed for changes and IL-1β blockadeconsistently led to a decrease in the numbers of neutrophils andgranulocytic MDSC in the tumors. Often these were accompanied bydecreased monocytes and monocytic MDSC; however, there was morevariability in these populations. Neutrophils both produce IL-1β andrespond to IL-1β while MDSC generation is often dependent on IL-1β, andboth subsets of cells can suppress the function of other immune cells.Decreases in both neutrophils and MDSC combined with a decrease in Tregsmay mean that the tumor microenvironment becomes less immune suppressivefollowing IL-1β blockade. A less suppressive TME may lead to a betteranti-tumor immune response, particularly with checkpoint blockade.

These data taken together show that blocking both IL-1β and thePD-1/PD-L1 axis may lead to a more immune active tumor microenvironmentor combining IL-1β blockade with chemotherapy may have a similar impact.

Example 10 Determining Immunogenicity/Allergenicity to IL-1β Antibody

During the CANTOS trial, blood samples for immunogenicity assessmentswere collected at baseline Month 12, 24 and end of study visit.Immunogenicity was analyzed using a bridging immunogenicityelectrochemiluminescence immunoassay (ECLIA). Samples were pre-treatedwith acetic acid and neutralized in buffer containing labeled drug(biotinylated ACZ885 and sulfo-TAG (Ruthenium) labeled ACZ885).Anti-canakinumab antibodies (anti-drug antibodies) were captured by acombination of biotinylated and sulfo-TAG labeled forms of ACZ885.

Complex formation was subsequently detected by electrochemiluminescenceby capturing complexes on Mesoscale Discovery Streptavidin (MSD) plates.

Treatment-emergent anticanakinumab antibodies (anti-drug antibodies)were detected in low and comparable proportions of patients across alltreatment groups (0.3%, 0.4% and 0.5% in the canakinumab 300 mg, 150 mgand placebo groups respectively) and were not associated withimmunogenicity related AEs or altered hsCRP response.

Example 11

Biomarker Analysis from the CANTOS Trial

Patients with gastric cancer, colorectal cancer and pancreatic cancerwere grouped into GI group. Patients with bladder cancer, renal cellcarcinoma and prostate cancer were grouped into GU group. Within thegroup, patients were further divided according to their baseline IL-6 orCRP level into above median group and below median group. The mean andmedian of time to cancer event were calculated as shown the table below.

There seems to have a trend that patient group have below median levelof CRP and IL-6 had in general longer time to develop cancer. This trendseems to be stronger based on IL-6 analysis than CRP, possibly due tothe fact that IL-6 is immediately downstream of IL-1b, where CTP isfurther away from IL-1b signaling and therefore could be influenced byother factors as well.

Time to cancer AE Set IL-6 median N Mean Median GI Above median 34 18.3516.03 Below median 35 27.84 28.55 GU Above median 33 21.79 17.45 Belowmedian 33 27.85 23.39

Time to cancer AE Set CRP median N Mean Median GI Above Median 56 19.2315.08 Below Median 58 25.61 26.17 GU Above Median 54 24.57 23.23 BelowMedian 56 25.15 24.13

Example 12

Randomized Phase II Study Assessing Efficacy and Safety ofCanakinumab+Asciminib Vs. Asciminib in CML Newly Diagnosed Patients orPatients who have Failed or are Intolerant to one Prior TKI

The study comprises a safety run-in part assessing the safety ofcanakinumab 200 mg subcutaneous (s.c.) every 28 days in combination withasciminib 40 mg BID (n=˜9 patients) followed by a randomized part (n=˜50patients) assessing canakinumab in combination with asciminib vs.asciminib. The primary objective is MR4.5 at 12 months and key secondaryobjective is TFR eligibility at 96 weeks of treatment.

Inclusion Criteria

-   -   Newly diagnosed CML-chronic phase or confirmed diagnosis of        CML-CP with resistance or intolerance to one prior TKI therapy.    -   Failure or intolerance to one prior TKI therapy at the time of        screening (adapted from the 2013 ELN Guidelines, Bacarrani 2013,        doi: 10.1182/blood-2013-05-501569). Failure is defined for        CML-CP patients (CP at the time of initiation of last therapy)        as follows (patients must meet at least one of the following        criteria):        -   Three months after the initiation of therapy: No CHR            (complete hematologic response) or >95% Ph⁺ metaphases.        -   Six months after the initiation of therapy: BCR-ABL            ratio >10% IS and/or >65% Ph⁺ metaphases.        -   Twelve months after initiation of therapy: BCR-ABL            ratio >10% IS and/or >35% Ph⁺ metaphases.        -   At any time after the initiation of therapy, loss of CHR,            CCyR (complete cytogenetic response) or PCyR (partial            cytogenetic response).        -   At any time after the initiation of therapy, the development            of new BCR-ABL mutations.        -   At any time after the initiation of therapy, confirmed loss            of MMR (major molecular response) in 2 consecutive tests, of            which one must have a BCR-ABL ratio ≥1% IS.        -   At any time after the initiation of therapy, new clonal            chromosome abnormalities in Ph⁺ cells: CCA/Ph+.    -   Intolerance is defined as:        -   Non-hematologic intolerance: Patients with grade 3 or 4            toxicity while on therapy, or with persistent grade 2            toxicity, unresponsive to optimal management, including dose            adjustments (unless dose reduction is not considered in the            best interest of the patient if response is already            suboptimal).        -   Hematologic intolerance: Patients with grade 3 or 4 toxicity            (absolute neutrophil count [ANC] or platelets) while on            therapy that is recurrent after dose reduction to the lowest            doses recommended by manufacturer.    -   Eastern Cooperative Oncology Group (ECOG) Performance Status        (PS) Score 0-1.    -   Adequate organ function.

Exclusion Criteria

-   -   Previous treatment with a hematopoietic stem-cell        transplantation or patient planning to undergo allogeneic        hematopoietic stem cell transplantation.    -   Cardiac or cardiac repolarization abnormality, including any of        the following:        -   History within 6 months prior to starting study treatment of            myocardial infarction (MI), angina pectoris, coronary artery            bypass graft (CABG).        -   Clinically significant cardiac arrhythmias (e.g.,            ventricular tachycardia), complete left bundle branch block,            high-grade AV block (e.g., bifascicular block, Mobitz type            II and third degree AV block).        -   QTcF at screening ≥450 ms (male patients), ≥460 ms (female            patients).        -   Long QT syndrome, family history of idiopathic sudden death            or congenital long QT syndrome, or any of the following:            -   Risk factors for Torsades de Pointes (TdP) including                uncorrected hypokalemia or hypomagnesemia, history of                cardiac failure, or history of clinically                significant/symptomatic bradycardia.            -   Concomitant medication(s) with a known risk to prolong                the QT interval and/or known to cause Torsades de                Pointes that cannot be discontinued or replaced 7 days                prior to starting study drug by safe alternative                medication.            -   Inability to determine the QTcF interval.    -   Severe and/or uncontrolled concurrent medical disease that in        the opinion of the investigator could cause unacceptable safety        risks or compromise compliance with the protocol (e.g.,        uncontrolled diabetes, active or uncontrolled infection,        pulmonary hypertension).    -   Adequate contraception.

1. An IL-1β binding antibody or a functional fragment thereof for use inthe treatment of CML in a patient.
 2. An IL-1β binding antibody or afunctional fragment thereof for use according to claim 1, wherein CMLhas at least a partial inflammatory basis.
 3. An IL-1β binding antibodyor a functional fragment thereof for use according to claim 1 or 2,wherein the IL-1β binding antibody or a functional fragment thereof iscanakinumab.
 4. An IL-1β binding antibody or a functional fragmentthereof for use according to claim 3, wherein the therapeutic effectiveamount of canakinumab is about 200 mg.
 5. An IL-1β binding antibody or afunctional fragment thereof for use according to claim 4, whereincanakinumab is administered about every 3 weeks or about every 4 weeks.6. An IL-1β binding antibody or a functional fragment thereof for useaccording to claim 3-5, wherein canakinumab is administeredsubcutaneously.
 7. An IL-1β binding antibody or a functional fragmentthereof for use according to claim 1 or 2, wherein the IL-1β bindingantibody or a functional fragment thereof is gevokizumab.
 8. An IL-1βbinding antibody or a functional fragment thereof for use according toclaim 7, wherein the therapeutic effective amount of gevokizumab isabout 30-120 mg.
 9. An IL-1β binding antibody or a functional fragmentthereof for use according to claim 8, wherein gevokizumab isadministered about every 3 weeks or about every 4 weeks.
 10. An IL-1βbinding antibody or a functional fragment thereof for use according toclaim 7-9, wherein gevokizumab is administered intravenously orsubcutaneously.
 11. An IL-1β binding antibody or a functional fragmentthereof for use according to any one of preceding claims, wherein atherapeutically effective amount of IL-1β binding antibody or afunctional fragment thereof is administered to the patient about every 3weeks or about every 4 weeks for at least about 13 months.
 12. An IL-1βbinding antibody or a functional fragment thereof for use according toany one of preceding claims, wherein the hazard rate of cancer mortalityof the patient is reduced by at least about 10%.
 13. An IL-1β bindingantibody or a functional fragment thereof for use according to any oneof preceding claims, wherein the patient has at least about 3 monthsprogression free survival (PFS).
 14. An IL-1β binding antibody or afunctional fragment thereof for use according to any one of precedingclaims, wherein the PFS of the patient is at least about 3 monthsprogression free survival (PFS) longer than standard of care treatment.15. An IL-1β binding antibody or a functional fragment thereof for useaccording to any one of preceding claims, wherein the patient has atleast about 3 months overall survival (OS).
 16. An IL-1β bindingantibody or a functional fragment thereof for use according to any oneof preceding claims, wherein the patient has at least about 3 monthsoverall survival (OS) longer than standard of care treatment.
 17. AnIL-1β binding antibody or a functional fragment thereof for useaccording to any one of preceding claims, wherein the patient is not athigh risk of developing serious infection.
 18. An IL-1β binding antibodyor a functional fragment thereof for use according to any one ofpreceding claims, wherein the IL-1β binding antibody or a functionalfragment thereof is not administered in combination with a TNFinhibitor.
 19. An IL-1β binding antibody or a functional fragmentthereof for use according to any one of preceding claims, wherein thepatient has at least 3 months disease free survival (DFS).
 20. An IL-1βbinding antibody or a functional fragment thereof for use according toany one of preceding claims, wherein the chance the patient developsantibody against said IL-1β binding antibody is less than about 1%. 21.An IL-1β binding antibody or a functional fragment thereof for useaccording to claim 20, wherein the IL-1β binding antibody or afunctional fragment thereof is canakinumab.
 22. An IL-1β bindingantibody or a functional fragment thereof for use according to any oneof preceding claims, wherein said patient has high sensitivityC-reactive protein (hsCRP) equal to or greater than about 2.1 mg/Lbefore first administration of said IL-1β binding antibody or functionalfragment thereof.
 23. An IL-1β binding antibody or a functional fragmentthereof for use according to any one of preceding claims, wherein saidIL-1β binding antibody or a functional fragment thereof is administeredin combination with one or more therapeutic agents.
 24. An IL-1β bindingantibody or a functional fragment thereof for use according to claim 23,wherein the one or more therapeutic agents is the standard of care agentfor CML.
 25. An IL-1β binding antibody or a functional fragment thereoffor use according to claim 23 or 24, wherein the one or more therapeuticagents is a checkpoint inhibitor.
 26. An IL-1β binding antibody or afunctional fragment thereof for use according to claim 25, wherein thecheckpoint inhibitor is selected from a list consisting of nivolumab,pembrolizumab, atezolizumab, durvalumab, avelumab, Ipilimumab andspartalizumab.
 27. An IL-1β binding antibody or a functional fragmentthereof for use according to claim 25, wherein the checkpoint inhibitoris pembrolizumab.
 28. An IL-1β binding antibody or a functional fragmentthereof for use according to any one of preceding claims, wherein saidIL-1β binding antibody or a functional fragment thereof is used, aloneor preferably in combination, as the first, second or third linetreatment.
 29. An IL-1β binding antibody or a functional fragmentthereof for use according to any one of preceding claims, wherein saidIL-1β binding antibody or a functional fragment thereof is used, aloneor preferably in combination, for more than one lines of treatment inthe same patient.
 30. An IL-1β binding antibody or a functional fragmentthereof for use according to any one claims 23, 24, 28-30, wherein saidone or more therapeutic agent is a BCR-ABL inhibitor.
 31. An IL-1βbinding antibody or a functional fragment thereof for use according toclaim 30, wherein said BCR-ABL inhibitor is selected from the groupconsisting of imatinib, nilotinib, dasatinib, dosutinib, radotinib,asciminib, ponatinib and bafetinib.
 32. An IL-1β binding antibody or afunctional fragment thereof for use according to claim 30, wherein saidBCR-ABL inhibitor is nilotinib.
 33. An IL-1β binding antibody or afunctional fragment thereof for use according to claim 30, wherein saidBCR-ABL inhibitor is asciminib.
 34. An IL-1β binding antibody or afunctional fragment thereof for use according to claim 33, whereinasciminib is administered at a dose of about 40 mg BID, in combinationwith a dose of canakinumab of about 200 mg intravenously about everyfour weeks (monthly).
 35. An IL-1β binding antibody or a functionalfragment thereof for use according to claim 34, wherein the patient isnewly diagnosed CML-chronic phase (CP).
 36. An IL-1β binding antibody ora functional fragment thereof for use according to claim 34, wherein thepatient is diagnosed with CML-CP with resistance, intolerance or failureto a prior TKI therapy.